LOCUS       V035155                 9439 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V035155
VERSION     V035155
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..3465
                     /label="MICB(NM_005931)"
                     /note="MICB(NM_005931)"
                     /gene="MICB"
     CDS             3472..3537
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3581..3792
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3805..4073
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4102..4111
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4132..4794
                     /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             4864..4917
                     /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             4918..5517
                     /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    5518..6106
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6180..6413
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6485..6606
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6646..6781
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6814..6830)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6838..6854
                     /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(6862..6892)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6907..6928
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7216..7804)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7975..8835)
                     /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(8836..8940)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9414..9430
                     /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 gccaccatgg ggctgggccg ggtcctgctg
     2341 tttctggccg tcgccttccc ttttgcaccc ccggcagccg ccgctgagcc ccacagtctt
     2401 cgttacaacc tcatggtgct gtcccaggat ggatctgtgc agtcagggtt tctcgctgag
     2461 ggacatctgg atggtcagcc cttcctgcgc tatgacaggc agaaacgcag ggcaaagccc
     2521 cagggacagt gggcagaaaa tgtcctggga gctaagacct gggacacaga gaccgaggac
     2581 ttgacagaga atgggcaaga cctcaggagg accctgactc atatcaagga ccagaaagga
     2641 ggcttgcatt ccctccagga gattagggtc tgtgagatcc atgaagacag cagcaccagg
     2701 ggctcccggc atttctacta cgatggggag ctcttcctct cccaaaacct ggagactcaa
     2761 gaatcgacag tgccccagtc ctccagagct cagaccttgg ctatgaacgt cacaaatttc
     2821 tggaaggaag atgccatgaa gaccaagaca cactatcgcg ctatgcaggc agactgcctg
     2881 cagaaactac agcgatatct gaaatccggg gtggccatca ggagaacagt gccccccatg
     2941 gtgaatgtca cctgcagcga ggtctcagag ggcaacatca ccgtgacatg cagggcttcc
     3001 agcttctatc cccggaatat cacactgacc tggcgtcagg atggggtatc tttgagccac
     3061 aacacccagc agtgggggga tgtcctgcct gatgggaatg gaacctacca gacctgggtg
     3121 gccaccagga ttcgccaagg agaggagcag aggttcacct gctacatgga acacagcggg
     3181 aatcacggca ctcaccctgt gccctctggg aaggcgctgg tgcttcagag tcaacggaca
     3241 gactttccat atgtttctgc tgctatgcca tgttttgtta ttattattat tctctgtgtc
     3301 ccttgttgca agaagaaaac atcagcggca gagggtccag agcttgtgag cctgcaggtc
     3361 ctggatcaac acccagttgg gacaggagac cacagggatg cagcacagct gggatttcag
     3421 cctctgatgt cagctactgg gtccactggt tccactgagg gcaccggatc cgactacaaa
     3481 gaccatgacg gtgattataa agatcatgac atcgactaca aggatgacga tgacaagtag
     3541 cggccgcgaa ggatctgcga tcgctccggt gcccgtcagt gggcagagcg cacatcgccc
     3601 acagtccccg agaagttggg gggaggggtc ggcaattgaa cgggtgccta gagaaggtgg
     3661 cgcggggtaa actgggaaag tgatgtcgtg tactggctcc gcctttttcc cgagggtggg
     3721 ggagaaccgt atataagtgc agtagtcgcc gtgaacgttc tttttcgcaa cgggtttgcc
     3781 gccagaacac agctgaagct tcgaggggct cgcatctctc cttcacgcgc ccgccgccct
     3841 acctgaggcc gccatccacg ccggttgagt cgcgttctgc cgcctcccgc ctgtggtgcc
     3901 tcctgaactg cgtccgccgt ctaggtaagt ttaaagctca ggtcgagacc gggcctttgt
     3961 ccggcgctcc cttggagcct acctagactc agccggctct ccacgctttg cctgaccctg
     4021 cttgctcaac tctacgtctt tgtttcgttt tctgttctgc gccgttacag atccaagctg
     4081 tgaccggcgc ctacgctaga cgccaccatg gagagcgacg agagcggcct gcccgccatg
     4141 gagatcgagt gccgcatcac cggcaccctg aacggcgtgg agttcgagct ggtgggcggc
     4201 ggagagggca cccccaagca gggccgcatg accaacaaga tgaagagcac caaaggcgcc
     4261 ctgaccttca gcccctacct gctgagccac gtgatgggct acggcttcta ccacttcggc
     4321 acctacccca gcggctacga gaaccccttc ctgcacgcca tcaacaacgg cggctacacc
     4381 aacacccgca tcgagaagta cgaggacggc ggcgtgctgc acgtgagctt cagctaccgc
     4441 tacgaggccg gccgcgtgat cggcgacttc aaggtggtgg gcaccggctt ccccgaggac
     4501 agcgtgatct tcaccgacaa gatcatccgc agcaacgcca ccgtggagca cctgcacccc
     4561 atgggcgata acgtgctggt gggcagcttc gcccgcacct tcagcctgcg cgacggcggc
     4621 tactacagct tcgtggtgga cagccacatg cacttcaaga gcgccatcca ccccagcatc
     4681 ctgcagaacg ggggccccat gttcgccttc cgccgcgtgg aggagctgca cagcaacacc
     4741 gagctgggca tcgtggagta ccagcacgcc ttcaagaccc ccatcgcctt cgccagatcc
     4801 cgcgctcagt cgtccaattc tgccgtggac ggcaccgccg gacccggctc caccggatct
     4861 cgcgagggca gaggaagtct tctaacatgc ggtgacgtgg aggagaatcc cggccctatg
     4921 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     4981 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5041 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5101 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5161 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5221 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5281 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5341 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5401 acctccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5461 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgaaat
     5521 caacctctgg attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct
     5581 tttacgctat gtggatacgc tgctttaatg cctttgtatc atgctattgc ttcccgtatg
     5641 gctttcattt tctcctcctt gtataaatcc tggttgctgt ctctttatga ggagttgtgg
     5701 cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt
     5761 tggggcattg ccaccacctg tcagctcctt tccgggactt tcgctttccc cctccctatt
     5821 gccacggcgg aactcatcgc cgcctgcctt gcccgctgct ggacaggggc tcggctgttg
     5881 ggcactgaca attccgtggt gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc
     5941 tgtgttgcca cctggattct gcgcgggacg tccttctgct acgtcccttc ggccctcaat
     6001 ccagcggacc ttccttcccg cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc
     6061 cttcgccctc agacgagtcg gatctccctt tgggccgcct ccccgcctgg tacctttaag
     6121 accaatgact tacaaggcag ctgtagatct tagccacttt ttaaaagaaa aggggggact
     6181 ggaagggcta attcactccc aacgaagata agatctgctt tttgcttgta ctgggtctct
     6241 ctggttagac cagatctgag cctgggagct ctctggctaa ctagggaacc cactgcttaa
     6301 gcctcaataa agcttgcctt gagtgcttca agtagtgtgt gcccgtctgt tgtgtgactc
     6361 tggtaactag agatccctca gaccctttta gtcagtgtgg aaaatctcta gcagtagtag
     6421 ttcatgtcat cttattattc agtatttata acttgcaaag aaatgaatat cagagagtga
     6481 gaggaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     6541 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     6601 atcttatcat gtctggctct agctatcccg cccctaactc cgcccatccc gcccctaact
     6661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     6721 gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     6781 ctagactttt gcagagacca aattcgtaat catgtcatag ctgtttcctg tgtgaaattg
     6841 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6901 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6961 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     7021 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     7081 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7141 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7201 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7261 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7321 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7381 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7441 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7501 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7561 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7621 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7681 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7741 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7801 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7861 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7921 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7981 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     8041 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     8101 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8161 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8221 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8281 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8341 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8401 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8461 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8521 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8581 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8641 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8701 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8761 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8821 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8881 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8941 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
     9001 ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga
     9061 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     9121 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     9181 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     9241 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
     9301 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
     9361 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
     9421 gacggccagt gccaagctg
//
