LOCUS       V027465                 9220 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V027465
VERSION     V027465
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..3246
                     /label="PSMD14(NM_005805)"
                     /note="PSMD14(NM_005805)"
                     /gene="PSMD14"
     CDS             3253..3318
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     promoter        3362..3573
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             3586..3854
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      3883..3892
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             3913..4575
                     /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             4645..4698
                     /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             4699..5298
                     /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    5299..5887
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5961..6194
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6266..6387
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6427..6562
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6595..6611)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6619..6635
                     /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(6643..6673)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6688..6709
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6997..7585)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7756..8616)
                     /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(8617..8721)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9195..9211
                     /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 acagacttct tagacttgga
     2341 ggaggtatgc ctggactggg ccaggggcca cctacagatg ctcctgcagt ggacacagca
     2401 gaacaagtct atatctcttc cctggcactg ttaaaaatgt taaaacatgg ccgtgctgga
     2461 gttccaatgg aagttatggg tttgatgctt ggagaatttg ttgatgatta taccgtcaga
     2521 gtgattgatg tgtttgctat gccacagtca ggaacaggtg tcagtgtgga ggcagttgat
     2581 ccagtgttcc aagctaaaat gttggatatg ttgaagcaga caggaaggcc ggagatggtt
     2641 gttggttggt atcacagtca ccctggcttt ggttgttggc tttctggtgt ggatatcaac
     2701 actcagcaga gctttgaagc cttgtcggag agagctgtgg cagtggttgt ggatcccatt
     2761 cagagtgtaa aaggaaaggt tgttattgat gccttcagat tgatcaatgc taatatgatg
     2821 gtcttaggac atgaaccaag acaaacaact tcgaatctgg gtcacttaaa caagccatct
     2881 atccaggcat taattcatgg actaaacaga cattattact ccattactat taactatcgg
     2941 aaaaatgaac tggaacagaa gatgttgcta aatttgcata agaagagttg gatggaaggt
     3001 ttgacacttc aggactacag tgaacattgt aaacacaatg aatcagtggt aaaagagatg
     3061 ttggaattag ccaagaatta caataaggct gtagaagaag aagataagat gacacctgaa
     3121 cagctggcaa taaagaatgt tggcaagcag gaccccaaac gtcatttgga ggaacatgtg
     3181 gatgtactta tgacctcaaa tattgtccag tgtttagcag ctatgttgga tactgtcgta
     3241 tttaaaggat ccgactacaa agaccatgac ggtgattata aagatcatga catcgactac
     3301 aaggatgacg atgacaagta gcggccgcga aggatctgcg atcgctccgg tgcccgtcag
     3361 tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt cggcaattga
     3421 acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt gtactggctc
     3481 cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc cgtgaacgtt
     3541 ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc tcgcatctct
     3601 ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag tcgcgttctg
     3661 ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag tttaaagctc
     3721 aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact cagccggctc
     3781 tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt ttctgttctg
     3841 cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat ggagagcgac
     3901 gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct gaacggcgtg
     3961 gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat gaccaacaag
     4021 atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca cgtgatgggc
     4081 tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt cctgcacgcc
     4141 atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg cggcgtgctg
     4201 cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt caaggtggtg
     4261 ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg cagcaacgcc
     4321 accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt cgcccgcacc
     4381 ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat gcacttcaag
     4441 agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt ccgccgcgtg
     4501 gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc cttcaagacc
     4561 cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga cggcaccgcc
     4621 ggacccggct ccaccggatc tcgcgagggc agaggaagtc ttctaacatg cggtgacgtg
     4681 gaggagaatc ccggccctat gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac
     4741 gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc
     4801 cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga actcttcctc
     4861 acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg
     4921 gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc
     4981 atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg cctcctggcg
     5041 ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac
     5101 cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc
     5161 ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc tccccttcta cgagcggctc
     5221 ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc
     5281 cgcaagcccg gtgcctgaaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     5341 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     5401 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     5461 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     5521 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     5581 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     5641 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaaatcatcg
     5701 tcctttcctt ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     5761 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     5821 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     5881 tccccgcctg gtacctttaa gaccaatgac ttacaaggca gctgtagatc ttagccactt
     5941 tttaaaagaa aaggggggac tggaagggct aattcactcc caacgaagat aagatctgct
     6001 ttttgcttgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     6061 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     6121 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     6181 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     6241 gaaatgaata tcagagagtg agaggaactt gtttattgca gcttataatg gttacaaata
     6301 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     6361 tttgtccaaa ctcatcaatg tatcttatca tgtctggctc tagctatccc gcccctaact
     6421 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     6481 atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag
     6541 tgaggaggct tttttggagg cctagacttt tgcagagacc aaattcgtaa tcatgtcata
     6601 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6661 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6721 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6781 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6841 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6901 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6961 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7021 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7081 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     7141 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     7201 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     7261 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7321 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7381 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7441 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7501 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7561 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7621 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7681 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7741 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7801 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7861 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7921 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7981 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8041 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     8101 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8161 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8221 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8281 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8341 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8401 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8461 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8521 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8581 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8641 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8701 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     8761 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc
     8821 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     8881 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     8941 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     9001 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcgccattc
     9061 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     9121 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     9181 ccagtcacga cgttgtaaaa cgacggccag tgccaagctg
//
