LOCUS       V029512                 8936 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029512
VERSION     V029512
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    1798..1915
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
                     /label="cPPT/CTS"
     promoter        2005..2208
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2297..3688
                     /label="CEP55(NM_001127182)"
                     /note="CEP55(NM_001127182)"
                     /gene="CEP55"
     promoter        3739..3950
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3963..4231
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4260..4269
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4290..4952
                     /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"
     misc_feature    5031..5619
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5693..5926
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5998..6119
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6138..6273
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6311..6327)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6335..6351
                     /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(6359..6389)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6404..6425
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6713..7301)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7472..8332)
                     /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(8333..8437)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8911..8927
                     /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 ggttaacttt
     1801 taaaagaaaa ggggggattg gggggtacag tgcaggggaa agaatagtag acataatagc
     1861 aacagacata caaactaaag aattacaaaa acaaattaca aaaattcaaa attttatcga
     1921 tactagtatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
     1981 gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga
     2041 tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg
     2101 ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg
     2161 caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac
     2221 cgtcagatcg cctggagacg ccatccacgc tgttttgacc tccatagaag attctagagc
     2281 tagcgaattc gccaccatgt cttccagaag taccaaagat ttaattaaaa gtaagtgggg
     2341 atcgaagcct agtaactcca aatccgaaac tacattagaa aaattaaagg gagaaattgc
     2401 acacttaaag acatcagtgg atgaaatcac aagtgggaaa ggaaagctga ctgataaaga
     2461 gagacacaga cttttggaga aaattcgagt ccttgaggct gagaaggaga agaatgctta
     2521 tcaactcaca gagaaggaca aagaaataca gcgactgaga gaccaactga aggccagata
     2581 tagtactacc acattgcttg aacagctgga agagacaacg agagaaggag aaaggaggga
     2641 gcaggtgttg aaagccttat ctgaagagaa agacgtattg aaacaacagt tgtctgctgc
     2701 aacctcacga attgctgaac ttgaaagcaa aaccaataca ctccgtttat cacagactgt
     2761 ggctccaaac tgcttcaact catcaataaa taatattcat gaaatggaaa tacagctgaa
     2821 agatgctctg gagaaaaatc agcagtggct cgtgtatgat cagcagcggg aagtctatgt
     2881 aaaaggactt ttagcaaaga tctttgagtt ggaaaagaaa acggaaacag ctgctcattc
     2941 actcccacag cagacaaaaa agcctgaatc agaaggttat cttcaagaag agaagcagaa
     3001 atgttacaac gatctcttgg caagtgcaaa aaaagatctt gaggttgaac gacaaaccat
     3061 aactcagctg agttttgaac tgagtgaatt tcgaagaaaa tatgaagaaa cccaaaaaga
     3121 agttcacaat ttaaatcagc tgttgtattc acaaagaagg gcagatgtgc aacatctgga
     3181 agatgatagg cataaaacag agaagataca aaaactcagg gaagagaatg atattgctag
     3241 gggaaaactt gaagaagaga agaagagatc cgaagagctc ttatctcagg tccagtttct
     3301 ttacacatct ctgctaaagc agcaagaaga acaaacaagg gtagctctgt tggaacaaca
     3361 gatgcaggca tgtactttag actttgaaaa tgaaaaactc gaccgtcaac atgtgcagca
     3421 tcaattgcat gtaattctta aggagctccg aaaagcaaga aatcaaataa cacagttgga
     3481 atccttgaaa cagcttcatg agtttgccat cacagagcca ttagtcactt tccaaggaga
     3541 gactgaaaac agagaaaaag ttgccgcctc accaaaaagt cccactgctg cactcaatga
     3601 aagcctggtg gaatgtccca agtgcaatat acagtatcca gccactgagc atcgcgatct
     3661 gcttgtccat gtggaatact gttcaaagta gggatccgcg gccgcgaagg atctgcgatc
     3721 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg
     3781 gaggggtcgg caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     3841 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     3901 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc
     3961 gaggggctcg catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc
     4021 ggttgagtcg cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct
     4081 aggtaagttt aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac
     4141 ctagactcag ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg
     4201 tttcgttttc tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg
     4261 ccaccatgga gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg
     4321 gcaccctgaa cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg
     4381 gccgcatgac caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc
     4441 tgagccacgt gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga
     4501 accccttcct gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg
     4561 aggacggcgg cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg
     4621 gcgacttcaa ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga
     4681 tcatccgcag caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg
     4741 gcagcttcgc ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca
     4801 gccacatgca cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt
     4861 tcgccttccg ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc
     4921 agcacgcctt caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg
     4981 ccgtggacgg caccgccgga cccggctcca ccggatctcg ctaagtcgac aatcaacctc
     5041 tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct ccttttacgc
     5101 tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt atggctttca
     5161 ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg tggcccgttg
     5221 tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact ggttggggca
     5281 ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct attgccacgg
     5341 cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg ttgggcactg
     5401 acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg
     5461 ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc aatccagcgg
     5521 accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc
     5581 ctcagacgag tcggatctcc ctttgggccg cctccccgcc tggtaccttt aagaccaatg
     5641 acttacaagg cagctgtaga tcttagccac tttttaaaag aaaagggggg actggaaggg
     5701 ctaattcact cccaacgaaa ataagatctg ctttttgctt gtactgggtc tctctggtta
     5761 gaccagatct gagcctggga gctctctggc taactaggga acccactgct taagcctcaa
     5821 taaagcttgc cttgagtgct tcaagtagtg tgtgcccgtc tgttgtgtga ctctggtaac
     5881 tagagatccc tcagaccctt ttagtcagtg tggaaaatct ctagcagtag tagttcatgt
     5941 catcttatta ttcagtattt ataacttgca aagaaatgaa tatcagagag tgagaggaac
     6001 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     6061 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     6121 catgtctggc tctagctatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6181 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     6241 tccagaagta gtgaggaggc ttttttggag gcctagactt ttgcagagac ggcccaaatt
     6301 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6361 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6421 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6481 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6541 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6601 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6661 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6721 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6781 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6841 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6901 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6961 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     7021 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     7081 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     7141 gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     7201 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7261 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7321 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7381 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7441 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7501 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7561 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7621 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7681 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7741 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7801 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7861 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7921 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7981 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     8041 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     8101 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     8161 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     8221 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8281 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8341 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8401 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8461 ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga
     8521 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     8581 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     8641 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     8701 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     8761 gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg
     8821 cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg
     8881 taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctg
//
