LOCUS       V038351                 9457 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038351
VERSION     V038351
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             2339..3745
                     /label="MMP1(NM_002421)"
                     /note="MMP1(NM_002421)"
                     /gene="MMP1"
     promoter        3810..4021
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4034..4302
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4331..4340
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4361..5023
                     /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             5093..5146
                     /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             5147..5545
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     misc_feature    5552..6140
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6214..6447
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6519..6640
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6659..6794
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6832..6848)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6856..6872
                     /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(6880..6910)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6925..6946
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7234..7822)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7993..8853)
                     /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(8854..8958)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9432..9448
                     /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 gtttacaatc gccaccctcg aggccaccat
     2341 gcacagcttt cctccactgc tgctgctgct gttctggggt gtggtgtctc acagcttccc
     2401 agcgactcta gaaacacaag agcaagatgt ggacttagtc cagaaatacc tggaaaaata
     2461 ctacaacctg aagaatgatg ggaggcaagt tgaaaagcgg agaaatagtg gcccagtggt
     2521 tgaaaaattg aagcaaatgc aggaattctt tgggctgaaa gtgactggga aaccagatgc
     2581 tgaaaccctg aaggtgatga agcagcccag atgtggagtg cctgatgtgg ctcagtttgt
     2641 cctcactgag gggaaccctc gctgggagca aacacatctg acctacagga ttgaaaatta
     2701 cacgccagat ttgccaagag cagatgtgga ccatgccatt gagaaagcct tccaactctg
     2761 gagtaatgtc acacctctga cattcaccaa ggtctctgag ggtcaagcag acatcatgat
     2821 atcttttgtc aggggagatc atcgggacaa ctctcctttt gatggacctg gaggaaatct
     2881 tgctcatgct tttcaaccag gcccaggtat tggaggggat gctcattttg atgaagatga
     2941 aaggtggacc aacaatttca gagagtacaa cttacatcgt gttgcagctc atgaactcgg
     3001 ccattctctt ggactctccc attctactga tatcggggct ttgatgtacc ctagctacac
     3061 cttcagtggt gatgttcagc tagctcagga tgacattgat ggcatccaag ccatatatgg
     3121 acgttcccaa aatcctgtcc agcccatcgg cccacaaacc ccaaaagcgt gtgacagtaa
     3181 gctaaccttt gatgctataa ctacgattcg gggagaagtg atgttcttta aagacagatt
     3241 ctacatgcgc acaaatccct tctacccgga agttgagctc aatttcattt ctgttttctg
     3301 gccacaactg ccaaatgggc ttgaagctgc ttacgaattt gccgacagag atgaagtccg
     3361 gtttttcaaa gggaataagt actgggctgt tcagggacag aatgtgctac acggataccc
     3421 caaggacatc tacagctcct ttggcttccc tagaactgtg aagcatatcg atgctgctct
     3481 ttctgaggaa aacactggaa aaacctactt ctttgttgct aacaaatact ggaggtatga
     3541 tgaatataaa cgatctatgg atccaggtta tcccaaaatg atagcacatg actttcctgg
     3601 aattggccac aaagttgatg cagttttcat gaaagatgga tttttctatt tctttcatgg
     3661 aacaagacaa tacaaatttg atcctaaaac gaagagaatt ttgactctcc agaaagctaa
     3721 tagctggttc aactgcagga aaaattaggg atccctgcga tatccagcgc ggccgcgaag
     3781 gatctgcgat cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga
     3841 gaagttgggg ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa
     3901 ctgggaaagt gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta
     3961 tataagtgca gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca
     4021 gctgaagctt cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg
     4081 ccatccacgc cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc
     4141 gtccgccgtc taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc
     4201 ttggagccta cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact
     4261 ctacgtcttt gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc
     4321 tacgctagac gccaccatgg agagcgacga gagcggcctg cccgccatgg agatcgagtg
     4381 ccgcatcacc ggcaccctga acggcgtgga gttcgagctg gtgggcggcg gagagggcac
     4441 ccccaagcag ggccgcatga ccaacaagat gaagagcacc aaaggcgccc tgaccttcag
     4501 cccctacctg ctgagccacg tgatgggcta cggcttctac cacttcggca cctaccccag
     4561 cggctacgag aaccccttcc tgcacgccat caacaacggc ggctacacca acacccgcat
     4621 cgagaagtac gaggacggcg gcgtgctgca cgtgagcttc agctaccgct acgaggccgg
     4681 ccgcgtgatc ggcgacttca aggtggtggg caccggcttc cccgaggaca gcgtgatctt
     4741 caccgacaag atcatccgca gcaacgccac cgtggagcac ctgcacccca tgggcgataa
     4801 cgtgctggtg ggcagcttcg cccgcacctt cagcctgcgc gacggcggct actacagctt
     4861 cgtggtggac agccacatgc acttcaagag cgccatccac cccagcatcc tgcagaacgg
     4921 gggccccatg ttcgccttcc gccgcgtgga ggagctgcac agcaacaccg agctgggcat
     4981 cgtggagtac cagcacgcct tcaagacccc catcgccttc gccagatccc gcgctcagtc
     5041 gtccaattct gccgtggacg gcaccgccgg acccggctcc accggatctc gcgagggcag
     5101 aggaagtctt ctaacatgcg gtgacgtgga ggagaatccc ggccctatgg ccaagccttt
     5161 gtctcaagaa gaatccaccc tcattgaaag agcaacggct acaatcaaca gcatccccat
     5221 ctctgaagac tacagcgtcg ccagcgcagc tctctctagc gacggccgca tcttcactgg
     5281 tgtcaatgta tatcatttta ctgggggacc ttgtgcagaa ctcgtggtgc tgggcactgc
     5341 tgctgctgcg gcagctggca acctgacttg tatcgtcgcg atcggaaatg agaacagggg
     5401 catcttgagc ccctgcggac ggtgccgaca ggtgcttctc gatctgcatc ctgggatcaa
     5461 agccatagtg aaggacagtg atggacagcc gacggcagtt gggattcgtg aattgctgcc
     5521 ctctggttat gtgtgggagg gctaagtcga caatcaacct ctggattaca aaatttgtga
     5581 aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat acgctgcttt
     5641 aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct ccttgtataa
     5701 atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt
     5761 gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca cctgtcagct
     5821 cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca tcgccgcctg
     5881 ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg tggtgttgtc
     5941 ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga ttctgcgcgg
     6001 gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt cccgcggcct
     6061 gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga gtcggatctc
     6121 cctttgggcc gcctccccgc ctggtacctt taagaccaat gacttacaag gcagctgtag
     6181 atcttagcca ctttttaaaa gaaaaggggg gactggaagg gctaattcac tcccaacgaa
     6241 aataagatct gctttttgct tgtactgggt ctctctggtt agaccagatc tgagcctggg
     6301 agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc
     6361 ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct
     6421 tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt
     6481 tataacttgc aaagaaatga atatcagaga gtgagaggaa cttgtttatt gcagcttata
     6541 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     6601 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg ctctagctat
     6661 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     6721 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     6781 cttttttgga ggcctagact tttgcagaga cggcccaaat tcgtaatcat ggtcatagct
     6841 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     6901 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     6961 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     7021 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     7081 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     7141 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     7201 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     7261 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     7321 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     7381 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     7441 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     7501 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     7561 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     7621 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaaggacagt
     7681 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     7741 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     7801 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     7861 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     7921 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     7981 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8041 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8101 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     8161 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8221 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8281 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8341 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8401 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8461 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8521 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8581 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8641 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8701 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8761 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8821 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     8881 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     8941 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat
     9001 tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg
     9061 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg
     9121 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     9181 gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat
     9241 gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gccattcgcc
     9301 attcaggctg cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca
     9361 gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca
     9421 gtcacgacgt tgtaaaacga cggccagtgc caagctg
//
