LOCUS       V039258                 9505 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V039258
VERSION     V039258
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             234..414
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
                     /label="5' LTR (truncated)"
     misc_feature    458..583
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1076..1309
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1493..1537
                     /codon_start=1
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /product="antigenic peptide corresponding to amino acids
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik
                     et al., 2013)"
                     /transl_table=1
                     /translation="KNEQELLELDKWASL"
                     /label="gp41 peptide"
     misc_feature    1804..1921
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2011..2214
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2317..3531
                     /label="CTH(NM_001902)"
                     /note="CTH(NM_001902)"
                     /gene="CTH"
     CDS             3538..3603
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3647..3858
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3871..4139
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4168..4177
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4198..4860
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             4930..4983
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             4984..5583
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     misc_feature    5584..6172
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6246..6479
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6551..6672
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6712..6847
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6880..6896)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6904..6920
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(6928..6958)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6973..6994
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7282..7870)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8041..8901)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(8902..9006)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9480..9496
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
ORIGIN
        1 acgcgtgtag tcttatgcaa tactcttgta gtcttgcaac atggtaacga tgagttagca
       61 acatgcctta caaggagaga aaaagcaccg tgcatgccga ttggtggaag taaggtggta
      121 cgatcgtgcc ttattaggaa ggcaacagac gggtctgaca tggattggac gaaccactga
      181 attgccgcat tgcagagata ttgtatttaa gtgcctagct cgatacaata aacgggtctc
      241 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      301 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      361 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
      421 cccgaacagg gacctgaaag cgaaagggaa accagagctc tctcgacgca ggactcggct
      481 tgctgaagcg cgcacggcaa gaggcgaggg gcggcgactg gtgagtacgc caaaaatttt
      541 gactagcgga ggctagaagg agagagatgg gtgcgagagc gtcagtatta agcgggggag
      601 aattagatcg cgatgggaaa aaattcggtt aaggccaggg ggaaagaaaa aatataaatt
      661 aaaacatata gtatgggcaa gcagggagct agaacgattc gcagttaatc ctggcctgtt
      721 agaaacatca gaaggctgta gacaaatact gggacagcta caaccatccc ttcagacagg
      781 atcagaagaa cttagatcat tatataatac agtagcaacc ctctattgtg tgcatcaaag
      841 gatagagata aaagacacca aggaagcttt agacaagata gaggaagagc aaaacaaaag
      901 taagaccacc gcacagcaag cggccactga tcttcagacc tggaggagga gatatgaggg
      961 acaattggag aagtgaatta tataaatata aagtagtaaa aattgaacca ttaggagtag
     1021 cacccaccaa ggcaaagaga agagtggtgc agagagaaaa aagagcagtg ggaataggag
     1081 ctttgttcct tgggttcttg ggagcagcag gaagcactat gggcgcagcc tcaatgacgc
     1141 tgacggtaca ggccagacaa ttattgtctg gtatagtgca gcagcagaac aatttgctga
     1201 gggctattga ggcgcaacag catctgttgc aactcacagt ctggggcatc aagcagctcc
     1261 aggcaagaat cctggctgtg gaaagatacc taaaggatca acagctcctg gggatttggg
     1321 gttgctctgg aaaactcatt tgcaccactg ctgtgccttg gaatgctagt tggagtaata
     1381 aatctctgga acagattgga atcacacgac ctggatggag tgggacagag aaattaacaa
     1441 ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag aaaagaatga
     1501 acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta acataacaaa
     1561 ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag gtttaagaat
     1621 agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac cattatcgtt
     1681 tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag aagaagaagg
     1741 tggagagaga gacagagaca gatccattcg attagtgaac ggatctcgac ggtatcggtt
     1801 aacttttaaa agaaaagggg ggattggggg gtacagtgca ggggaaagaa tagtagacat
     1861 aatagcaaca gacatacaaa ctaaagaatt acaaaaacaa attacaaaaa ttcaaaattt
     1921 tatcgatact agtattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     1981 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2041 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2101 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2161 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggttt atataagcag agctcgttta
     2221 gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagattc
     2281 tagagctagc ctgatacgaa ctcggaattc gccaccatgc aggaaaaaga cgcctcctca
     2341 caaggtttcc tgccacactt ccaacatttc gccacgcagg cgatccatgt gggccaggat
     2401 ccagagcaat ggacctccag ggctgtagtg ccccccatct cactgtccac cacgttcaag
     2461 caaggggcgc ctggccagca ctcgggtttt gaatatagcc gttctggaaa tcccactagg
     2521 aattgccttg aaaaagcagt ggcagcactg gatggggcta agtactgttt ggcctttgct
     2581 tcaggtttag cagccactgt aactattacc catcttttaa aagcaggaga ccaaattatt
     2641 tgtatggatg atgtgtatgg aggtacaaac aggtacttca ggcaagtggc atctgaattt
     2701 ggattaaaga tttcttttgt tgattgttcc aaaatcaaat tactagaggc agcaattaca
     2761 ccagaaacca agcttgtttg gatcgaaacc cccacaaacc ccacccagaa ggtgattgac
     2821 attgaaggct gtgcacatat tgtccataag catggagaca ttattttggt cgtggataac
     2881 acttttatgt caccatattt ccagcgccct ttggctctgg gagctgatat ttctatgtat
     2941 tctgcaacaa aatacatgaa tggccacagt gatgttgtaa tgggcctggt gtctgttaat
     3001 tgtgaaagcc ttcataatag acttcgtttc ttgcaaaact ctcttggagc agttccatct
     3061 cctattgatt gttacctctg caatcgaggt ctgaagactc tacatgtccg aatggaaaag
     3121 catttcaaaa acggaatggc agttgcccag ttcctggaat ctaatccttg ggtagaaaag
     3181 gttatttatc ctgggctgcc ctctcatcca cagcatgagt tggtgaagcg tcagtgtaca
     3241 ggttgtacag ggatggtcac cttttatatt aagggcactc ttcagcatgc tgagattttc
     3301 ctcaagaacc taaagctatt tactctggcc gagagcttgg gaggattcga aagccttgct
     3361 gagcttccgg caatcatgac tcatgcatca gttcttaaga atgacagaga tgtccttgga
     3421 attagtgaca cactgattcg actttctgtg ggcttagagg atgaggaaga cctactggaa
     3481 gatctagatc aagctttgaa ggcagcacac cctccaagtg gaagtcacag cggatccgac
     3541 tacaaagacc atgacggtga ttataaagat catgacatcg actacaagga tgacgatgac
     3601 aagtagcggc cgcgaaggat ctgcgatcgc tccggtgccc gtcagtgggc agagcgcaca
     3661 tcgcccacag tccccgagaa gttgggggga ggggtcggca attgaacggg tgcctagaga
     3721 aggtggcgcg gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag
     3781 ggtgggggag aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg
     3841 tttgccgcca gaacacagct gaagcttcga ggggctcgca tctctccttc acgcgcccgc
     3901 cgccctacct gaggccgcca tccacgccgg ttgagtcgcg ttctgccgcc tcccgcctgt
     3961 ggtgcctcct gaactgcgtc cgccgtctag gtaagtttaa agctcaggtc gagaccgggc
     4021 ctttgtccgg cgctcccttg gagcctacct agactcagcc ggctctccac gctttgcctg
     4081 accctgcttg ctcaactcta cgtctttgtt tcgttttctg ttctgcgccg ttacagatcc
     4141 aagctgtgac cggcgcctac gctagacgcc accatggaga gcgacgagag cggcctgccc
     4201 gccatggaga tcgagtgccg catcaccggc accctgaacg gcgtggagtt cgagctggtg
     4261 ggcggcggag agggcacccc caagcagggc cgcatgacca acaagatgaa gagcaccaaa
     4321 ggcgccctga ccttcagccc ctacctgctg agccacgtga tgggctacgg cttctaccac
     4381 ttcggcacct accccagcgg ctacgagaac cccttcctgc acgccatcaa caacggcggc
     4441 tacaccaaca cccgcatcga gaagtacgag gacggcggcg tgctgcacgt gagcttcagc
     4501 taccgctacg aggccggccg cgtgatcggc gacttcaagg tggtgggcac cggcttcccc
     4561 gaggacagcg tgatcttcac cgacaagatc atccgcagca acgccaccgt ggagcacctg
     4621 caccccatgg gcgataacgt gctggtgggc agcttcgccc gcaccttcag cctgcgcgac
     4681 ggcggctact acagcttcgt ggtggacagc cacatgcact tcaagagcgc catccacccc
     4741 agcatcctgc agaacggggg ccccatgttc gccttccgcc gcgtggagga gctgcacagc
     4801 aacaccgagc tgggcatcgt ggagtaccag cacgccttca agacccccat cgccttcgcc
     4861 agatcccgcg ctcagtcgtc caattctgcc gtggacggca ccgccggacc cggctccacc
     4921 ggatctcgcg agggcagagg aagtcttcta acatgcggtg acgtggagga gaatcccggc
     4981 cctatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     5041 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     5101 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     5161 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     5221 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     5281 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     5341 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     5401 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     5461 ctggagacct ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     5521 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     5581 tgaaatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     5641 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     5701 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     5761 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     5821 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     5881 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     5941 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     6001 ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     6061 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     6121 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc
     6181 tttaagacca atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg
     6241 gggactggaa gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg
     6301 gtctctctgg ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact
     6361 gcttaagcct caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg
     6421 tgactctggt aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag
     6481 tagtagttca tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga
     6541 gagtgagagg aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     6601 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     6661 caatgtatct tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc
     6721 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     6781 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     6841 ggaggcctag acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg
     6901 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6961 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     7021 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     7081 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     7141 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     7201 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     7261 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     7321 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     7381 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     7441 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     7501 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     7561 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     7621 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     7681 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     7741 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     7801 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     7861 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     7921 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     7981 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     8041 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8101 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8161 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     8221 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8281 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8341 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8401 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8461 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8521 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8581 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8641 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8701 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8761 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8821 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8881 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     8941 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     9001 tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac
     9061 attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga
     9121 cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc tgtaagcgga
     9181 tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg
     9241 gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     9301 accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat tcaggctgcg
     9361 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
     9421 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
     9481 taaaacgacg gccagtgcca agctg
//
