LOCUS       V031810                 9430 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031810
VERSION     V031810
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..3456
                     /label="PSMD4(NM_001330692)"
                     /note="PSMD4(NM_001330692)"
                     /gene="PSMD4"
     CDS             3463..3528
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3572..3783
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3796..4064
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4093..4102
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4123..4785
                     /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             4855..4908
                     /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             4909..5508
                     /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    5509..6097
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6171..6404
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6476..6597
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6637..6772
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6805..6821)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6829..6845
                     /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(6853..6883)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6898..6919
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7207..7795)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7966..8826)
                     /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(8827..8931)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9405..9421
                     /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 tgttggaaag cactatggtg
     2341 tgtgtggaca acagtgagta tatgcggaat ggagacttct tacccaccag gctgcaggcc
     2401 cagcaggatg ctgtcaacat agtttgtcat tcaaagaccc gcagcaaccc tgagaacaac
     2461 gtgggcctta tcacactggc taatgactgt gaagtgctga ccacactcac cccagacact
     2521 ggccgtatcc tgtccaagct acatactgtc caacccaagg gcaagatcac cttctgcacg
     2581 ggcatccgcg tggcccatct ggctctgaag caccgacaag gcaagaatca caagatgcgc
     2641 atcattgcct ttgtgggaag cccagtggag gacaatgaga aggatctggt gaaactggct
     2701 aaacgcctca agaaggagaa agtaaatgtt gacattatca attttgggga agaggaggtg
     2761 aacacagaaa agctgacagc ctttgtaaac acgttgaatg gcaaagatgg aaccggttct
     2821 catctggtga cagtgcctcc tgggcccagt ttggctgatg ctctcatcag ttctccgatt
     2881 ttggctggtg aaggtggtgc catgctgggt cttggtgcca gtgactttga atttggagta
     2941 gatcccagtg ctgatcctga gctggccttg gcccttcgtg tatctatgga agagcagcgg
     3001 cagcggcagg aggaggaggc ccggcgggca gctgcagctt ctgctgctga ggccgggatt
     3061 gctacgactg ggactgaagg tgaaagagac tcagacgatg ccctgctgaa gatgaccatc
     3121 agccagcaag agtttggccg cactgggctt cctgacctaa gcagtatgac tgaggaagag
     3181 cagattgctt atgccatgca gatgtccctg cagggagcag agtttggcca ggcggaatca
     3241 gcagacattg atgccagctc agctatggac acatctgagc cagccaagga ggaggatgat
     3301 tacgacgtga tgcaggaccc cgagttcctt cagagtgtcc tagagaacct cccaggtgtg
     3361 gatcccaaca atgaagccat tcgaaatgct atgggctccc tggcctccca ggccaccaag
     3421 gacggcaaga aggacaagaa ggaggaagac aagaagggat ccgactacaa agaccatgac
     3481 ggtgattata aagatcatga catcgactac aaggatgacg atgacaagta gcggccgcga
     3541 aggatctgcg atcgctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc
     3601 gagaagttgg ggggaggggt cggcaattga acgggtgcct agagaaggtg gcgcggggta
     3661 aactgggaaa gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg
     3721 tatataagtg cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca
     3781 cagctgaagc ttcgaggggc tcgcatctct ccttcacgcg cccgccgccc tacctgaggc
     3841 cgccatccac gccggttgag tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact
     3901 gcgtccgccg tctaggtaag tttaaagctc aggtcgagac cgggcctttg tccggcgctc
     3961 ccttggagcc tacctagact cagccggctc tccacgcttt gcctgaccct gcttgctcaa
     4021 ctctacgtct ttgtttcgtt ttctgttctg cgccgttaca gatccaagct gtgaccggcg
     4081 cctacgctag acgccaccat ggagagcgac gagagcggcc tgcccgccat ggagatcgag
     4141 tgccgcatca ccggcaccct gaacggcgtg gagttcgagc tggtgggcgg cggagagggc
     4201 acccccaagc agggccgcat gaccaacaag atgaagagca ccaaaggcgc cctgaccttc
     4261 agcccctacc tgctgagcca cgtgatgggc tacggcttct accacttcgg cacctacccc
     4321 agcggctacg agaacccctt cctgcacgcc atcaacaacg gcggctacac caacacccgc
     4381 atcgagaagt acgaggacgg cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc
     4441 ggccgcgtga tcggcgactt caaggtggtg ggcaccggct tccccgagga cagcgtgatc
     4501 ttcaccgaca agatcatccg cagcaacgcc accgtggagc acctgcaccc catgggcgat
     4561 aacgtgctgg tgggcagctt cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc
     4621 ttcgtggtgg acagccacat gcacttcaag agcgccatcc accccagcat cctgcagaac
     4681 gggggcccca tgttcgcctt ccgccgcgtg gaggagctgc acagcaacac cgagctgggc
     4741 atcgtggagt accagcacgc cttcaagacc cccatcgcct tcgccagatc ccgcgctcag
     4801 tcgtccaatt ctgccgtgga cggcaccgcc ggacccggct ccaccggatc tcgcgagggc
     4861 agaggaagtc ttctaacatg cggtgacgtg gaggagaatc ccggccctat gaccgagtac
     4921 aagcccacgg tgcgcctcgc cacccgcgac gacgtcccca gggccgtacg caccctcgcc
     4981 gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg atccggaccg ccacatcgag
     5041 cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg
     5101 tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg
     5161 ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc
     5221 gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc
     5281 ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg
     5341 ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg
     5401 ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg
     5461 cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgaaa tcaacctctg
     5521 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     5581 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     5641 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     5701 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     5761 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     5821 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     5881 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     5941 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6001 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6061 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gtacctttaa gaccaatgac
     6121 ttacaaggca gctgtagatc ttagccactt tttaaaagaa aaggggggac tggaagggct
     6181 aattcactcc caacgaagat aagatctgct ttttgcttgt actgggtctc tctggttaga
     6241 ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata
     6301 aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta
     6361 gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtagta gttcatgtca
     6421 tcttattatt cagtatttat aacttgcaaa gaaatgaata tcagagagtg agaggaactt
     6481 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6541 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     6601 tgtctggctc tagctatccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt
     6661 tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc
     6721 gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt
     6781 tgcagagacc aaattcgtaa tcatgtcata gctgtttcct gtgtgaaatt gttatccgct
     6841 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     6901 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     6961 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     7021 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     7081 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     7141 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     7201 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     7261 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     7321 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     7381 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     7441 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     7501 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     7561 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     7621 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     7681 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     7741 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     7801 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     7861 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     7921 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     7981 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8041 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     8101 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     8161 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8221 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     8281 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8341 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8401 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     8461 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     8521 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     8581 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     8641 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     8701 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     8761 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     8821 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     8881 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     8941 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     9001 taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     9061 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     9121 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     9181 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     9241 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     9301 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     9361 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     9421 tgccaagctg
//
