LOCUS       V028371                 8615 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028371
VERSION     V028371
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..3373
                     /label="ST8SIA4(NM_005668)"
                     /note="ST8SIA4(NM_005668)"
                     /gene="ST8SIA4"
     promoter        3418..3629
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3642..3910
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3939..3948
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3969..4631
                     /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    4710..5298
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5372..5605
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    5677..5798
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      5817..5952
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(5990..6006)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6014..6030
                     /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(6038..6068)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6083..6104
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6392..6980)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7151..8011)
                     /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(8012..8116)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     8590..8606
                     /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 gccaccatgc gctccattag gaagaggtgg acgatctgca caataagtct
     2341 gctcctgatc ttttataaga caaaagaaat agcaagaact gaggagcacc aggagacgca
     2401 actcatcgga gatggtgaat tgtctttgag tcggtcactt gtcaatagct ctgataaaat
     2461 cattcgaaag gctggctctt caatcttcca gcacaatgta gaaggttgga aaatcaattc
     2521 ctctttggtc ctagagataa ggaagaacat acttcgtttc ttagatgcag aacgagatgt
     2581 gtcagtggtc aagagcagtt ttaagcctgg tgatgtcata cactatgtgc ttgacaggcg
     2641 ccggacacta aacatttctc atgatctaca tagcctccta cctgaagttt caccaatgaa
     2701 gaatcgcagg tttaagacct gtgcagttgt tggaaattct ggcattctgt tagacagtga
     2761 atgtggaaag gagattgaca gtcacaattt tgtaataagg tgtaatctag ctcctgtggt
     2821 ggagtttgct gcagatgtgg gaactaaatc agattttatt accatgaatc catcagttgt
     2881 acaaagagca tttggaggct ttcgaaatga gagtgacaga gaaaaatttg tgcatagact
     2941 ttccatgctg aatgacagtg tcctttggat tcctgctttc atggtcaaag gaggagagaa
     3001 gcacgtggag tgggttaatg cattaatcct taagaataaa ctgaaagtgc gaactgccta
     3061 tccgtcattg agacttattc atgctgtcag aggttactgg ctgaccaaca aagttcctat
     3121 caaaagaccc agcacaggtc ttctcatgta tacacttgcc acaagattct gtgatgaaat
     3181 tcacctgtat ggattctggc ccttccctaa ggatttaaat ggaaaagcgg tcaaatatca
     3241 ttattatgat gacttaaaat ataggtactt ttccaatgca agccctcaca gaatgccatt
     3301 agaattcaaa acattaaatg tgctacataa tagaggagct ctaaaactga caacaggaaa
     3361 gtgtgtaaag caataagcgg ccgcgaagga tctgcgatcg ctccggtgcc cgtcagtggg
     3421 cagagcgcac atcgcccaca gtccccgaga agttgggggg aggggtcggc aattgaacgg
     3481 gtgcctagag aaggtggcgc ggggtaaact gggaaagtga tgtcgtgtac tggctccgcc
     3541 tttttcccga gggtggggga gaaccgtata taagtgcagt agtcgccgtg aacgttcttt
     3601 ttcgcaacgg gtttgccgcc agaacacagc tgaagcttcg aggggctcgc atctctcctt
     3661 cacgcgcccg ccgccctacc tgaggccgcc atccacgccg gttgagtcgc gttctgccgc
     3721 ctcccgcctg tggtgcctcc tgaactgcgt ccgccgtcta ggtaagttta aagctcaggt
     3781 cgagaccggg cctttgtccg gcgctccctt ggagcctacc tagactcagc cggctctcca
     3841 cgctttgcct gaccctgctt gctcaactct acgtctttgt ttcgttttct gttctgcgcc
     3901 gttacagatc caagctgtga ccggcgccta cgctagacgc caccatggag agcgacgaga
     3961 gcggcctgcc cgccatggag atcgagtgcc gcatcaccgg caccctgaac ggcgtggagt
     4021 tcgagctggt gggcggcgga gagggcaccc ccaagcaggg ccgcatgacc aacaagatga
     4081 agagcaccaa aggcgccctg accttcagcc cctacctgct gagccacgtg atgggctacg
     4141 gcttctacca cttcggcacc taccccagcg gctacgagaa ccccttcctg cacgccatca
     4201 acaacggcgg ctacaccaac acccgcatcg agaagtacga ggacggcggc gtgctgcacg
     4261 tgagcttcag ctaccgctac gaggccggcc gcgtgatcgg cgacttcaag gtggtgggca
     4321 ccggcttccc cgaggacagc gtgatcttca ccgacaagat catccgcagc aacgccaccg
     4381 tggagcacct gcaccccatg ggcgataacg tgctggtggg cagcttcgcc cgcaccttca
     4441 gcctgcgcga cggcggctac tacagcttcg tggtggacag ccacatgcac ttcaagagcg
     4501 ccatccaccc cagcatcctg cagaacgggg gccccatgtt cgccttccgc cgcgtggagg
     4561 agctgcacag caacaccgag ctgggcatcg tggagtacca gcacgccttc aagaccccca
     4621 tcgccttcgc cagatcccgc gctcagtcgt ccaattctgc cgtggacggc accgccggac
     4681 ccggctccac cggatctcgc taagtcgaca atcaacctct ggattacaaa atttgtgaaa
     4741 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     4801 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     4861 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     4921 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     4981 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     5041 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     5101 ggaaatcatc gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     5161 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     5221 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     5281 tttgggccgc ctccccgcct ggtaccttta agaccaatga cttacaaggc agctgtagat
     5341 cttagccact ttttaaaaga aaagggggga ctggaagggc taattcactc ccaacgaaaa
     5401 taagatctgc tttttgcttg tactgggtct ctctggttag accagatctg agcctgggag
     5461 ctctctggct aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt
     5521 caagtagtgt gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt
     5581 tagtcagtgt ggaaaatctc tagcagtagt agttcatgtc atcttattat tcagtattta
     5641 taacttgcaa agaaatgaat atcagagagt gagaggaact tgtttattgc agcttataat
     5701 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     5761 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggct ctagctatcc
     5821 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     5881 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     5941 tttttggagg cctagacttt tgcagagacg gcccaaattc gtaatcatgg tcatagctgt
     6001 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6061 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6121 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6181 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6241 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6301 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6361 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6421 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6481 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6541 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6601 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6661 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6721 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6781 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     6841 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6901 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6961 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     7021 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     7081 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     7141 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     7201 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     7261 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     7321 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     7381 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     7441 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7501 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7561 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7621 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7681 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7741 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7801 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7861 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7921 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7981 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     8041 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     8101 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     8161 ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt
     8221 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     8281 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     8341 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     8401 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgc cattcgccat
     8461 tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc
     8521 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
     8581 cacgacgttg taaaacgacg gccagtgcca agctg
//
