LOCUS       V042513                 9605 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V042513
VERSION     V042513
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..4213
                     /label="HSPA2(NM_001387931)"
                     /note="HSPA2(NM_001387931)"
                     /gene="HSPA2"
     CDS             4238..4282
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic
                     ribonuclease A"
                     /transl_table=1
                     /translation="KETAAAKFERQHMDS"
                     /label="S-Tag"
     CDS             4316..4339
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        4408..4619
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4632..4900
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4929..4938
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4959..5621
                     /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    5700..6288
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6362..6595
                     /note="self-inactivating&nbsp;3' long terminal repeat (LTR)
                     from HIV-1"
                     /label="3' LTR (ΔU3)"
     polyA_signal    6667..6788
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     rep_origin      6807..6942
                     /note="SV40 origin of replication"
                     /label="SV40 ori"
     primer_bind     complement(6980..6996)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7004..7020
                     /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(7028..7058)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7073..7094
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7382..7970)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8141..9001)
                     /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(9002..9106)
                     /gene="bla"
                     /label="AmpR promoter"
     primer_bind     9580..9596
                     /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 ctgcccgtgg cccggctatc ggcatcgacc tgggcaccac
     2341 ctattcgtgc gtcggggtct tccaacatgg caaggtggag atcatcgcca acgaccaggg
     2401 caatcgcacc acccccagct acgtggcctt cacggacacc gagcgcctca tcggcgacgc
     2461 cgccaagaac caggtggcca tgaaccccac caacaccatc ttcgacgcca agaggctgat
     2521 tggacggaaa ttcgaggatg ccacagtgca gtcggatatg aaacactggc cgttccgggt
     2581 ggtgagcgag ggaggcaagc ccaaagtgca agtagagtac aagggggaga ccaagacctt
     2641 cttcccagag gagatatcct ccatggtcct cacgaagatg aaggagatcg cggaagccta
     2701 cctggggggc aaggtgcaca gcgcggtcat aacggtcccg gcctatttca acgactcgca
     2761 gcgccaggcc accaaggacg caggcaccat cacggggctc aatgtgctgc gcatcatcaa
     2821 cgagcccacg gcggcggcca tcgcctacgg cctggacaag aagggctgcg cgggcggcga
     2881 gaagaacgtg ctcatctttg acctgggcgg tggcactttc gacgtgtcca tcctgaccat
     2941 cgaggatggc atcttcgagg tgaagtccac ggccggcgac acccacctgg gcggtgagga
     3001 cttcgacaac cgcatggtga gccacctggc ggaggagttc aagcgcaagc acaagaagga
     3061 cattgggccc aacaagcgcg ccgtgaggcg gctgcgcacc gcttgcgagc gcgccaagcg
     3121 caccctgagc tcgtccacgc aggcgagcat cgagatcgac tcgctctacg agggcgtgga
     3181 cttctatacg tccatcacgc gcgcccgctt cgaggagctc aatgccgacc tctttcgcgg
     3241 gaccctggag ccggtggaga aggcgctgcg cgacgccaag ctggacaagg gccagatcca
     3301 ggagatcgtg ctggtgggcg gctccactcg tatccccaag atccagaagc tgctgcagga
     3361 tttcttcaac ggcaaggagc tgaacaagag catcaacccc gacgaggcgg tggcctatgg
     3421 cgccgcggtg caggcggcca tcctcatcgg cgacaaatca gagaatgtgc aggacctgct
     3481 gctactcgac gtgaccccgt tgtcgctggg catcgagaca gctggcggtg tcatgacccc
     3541 actcatcaag aggaacacca cgatccccac caagcagacg cagaccttca ccacctactc
     3601 ggacaaccag agcagcgtac tggtgcaggt atacgagggc gaacgggcca tgaccaagga
     3661 caataacctg ctgggcaagt tcgacctgac cgggattccc cctgcgcctc gcggggtccc
     3721 ccaaatcgag gttaccttcg acattgacgc caatggcatc cttaacgtta ccgccgccga
     3781 caagagcacc ggtaaggaaa acaaaatcac catcaccaat gacaaaggtc gtctgagcaa
     3841 ggacgacatt gaccggatgg tgcaggaggc ggagcggtac aaatcggaag atgaggcgaa
     3901 tcgcgaccga gtcgcggcca aaaacgccct ggagtcctat acctacaaca tcaagcagac
     3961 ggtggaagac gagaaactga ggggcaagat tagcgagcag gacaaaaaca agatcctcga
     4021 caagtgtcag gaggtgatca actggctcga ccgaaaccag atggcagaga aagatgagta
     4081 tgaacacaag cagaaagagc tcgaaagagt ttgcaacccc atcatcagca aactttacca
     4141 aggtggtcct ggcggcggca gcggcggcgg cggttcagga gcctccgggg gacccaccat
     4201 cgaagaagtg gacggctcga gctcaagctt cgaattcaaa gaaaccgctg ctgctaaatt
     4261 cgaacgccag cacatggaca gcggagccgg cgcaggagcc ggcgctgacg cgcctgacta
     4321 caaagacgat gacgacaagg gagattacaa ggatgactag ggatccgcgg ccgcgaagga
     4381 tctgcgatcg ctccggtgcc cgtcagtggg cagagcgcac atcgcccaca gtccccgaga
     4441 agttgggggg aggggtcggc aattgaacgg gtgcctagag aaggtggcgc ggggtaaact
     4501 gggaaagtga tgtcgtgtac tggctccgcc tttttcccga gggtggggga gaaccgtata
     4561 taagtgcagt agtcgccgtg aacgttcttt ttcgcaacgg gtttgccgcc agaacacagc
     4621 tgaagcttcg aggggctcgc atctctcctt cacgcgcccg ccgccctacc tgaggccgcc
     4681 atccacgccg gttgagtcgc gttctgccgc ctcccgcctg tggtgcctcc tgaactgcgt
     4741 ccgccgtcta ggtaagttta aagctcaggt cgagaccggg cctttgtccg gcgctccctt
     4801 ggagcctacc tagactcagc cggctctcca cgctttgcct gaccctgctt gctcaactct
     4861 acgtctttgt ttcgttttct gttctgcgcc gttacagatc caagctgtga ccggcgccta
     4921 cgctagacgc caccatggag agcgacgaga gcggcctgcc cgccatggag atcgagtgcc
     4981 gcatcaccgg caccctgaac ggcgtggagt tcgagctggt gggcggcgga gagggcaccc
     5041 ccaagcaggg ccgcatgacc aacaagatga agagcaccaa aggcgccctg accttcagcc
     5101 cctacctgct gagccacgtg atgggctacg gcttctacca cttcggcacc taccccagcg
     5161 gctacgagaa ccccttcctg cacgccatca acaacggcgg ctacaccaac acccgcatcg
     5221 agaagtacga ggacggcggc gtgctgcacg tgagcttcag ctaccgctac gaggccggcc
     5281 gcgtgatcgg cgacttcaag gtggtgggca ccggcttccc cgaggacagc gtgatcttca
     5341 ccgacaagat catccgcagc aacgccaccg tggagcacct gcaccccatg ggcgataacg
     5401 tgctggtggg cagcttcgcc cgcaccttca gcctgcgcga cggcggctac tacagcttcg
     5461 tggtggacag ccacatgcac ttcaagagcg ccatccaccc cagcatcctg cagaacgggg
     5521 gccccatgtt cgccttccgc cgcgtggagg agctgcacag caacaccgag ctgggcatcg
     5581 tggagtacca gcacgccttc aagaccccca tcgccttcgc cagatcccgc gctcagtcgt
     5641 ccaattctgc cgtggacggc accgccggac ccggctccac cggatctcgc taagtcgaca
     5701 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     5761 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     5821 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     5881 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     5941 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     6001 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     6061 tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct tggctgctcg
     6121 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6181 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6241 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggtaccttta
     6301 agaccaatga cttacaaggc agctgtagat cttagccact ttttaaaaga aaagggggga
     6361 ctggaagggc taattcactc ccaacgaaaa taagatctgc tttttgcttg tactgggtct
     6421 ctctggttag accagatctg agcctgggag ctctctggct aactagggaa cccactgctt
     6481 aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt gtgcccgtct gttgtgtgac
     6541 tctggtaact agagatccct cagacccttt tagtcagtgt ggaaaatctc tagcagtagt
     6601 agttcatgtc atcttattat tcagtattta taacttgcaa agaaatgaat atcagagagt
     6661 gagaggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6781 gtatcttatc atgtctggct ctagctatcc cgcccctaac tccgcccagt tccgcccatt
     6841 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     6901 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctagacttt tgcagagacg
     6961 gcccaaattc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     7021 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     7081 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     7141 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     7201 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     7261 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     7321 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     7381 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     7441 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     7501 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     7561 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     7621 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     7681 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     7741 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     7801 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     7861 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     7921 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     7981 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     8041 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     8101 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     8161 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     8221 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     8281 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     8341 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     8401 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     8461 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     8521 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     8581 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     8641 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     8701 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     8761 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     8821 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     8881 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     8941 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     9001 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     9061 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     9121 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     9181 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     9241 tgcagctccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     9301 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     9361 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     9421 gaaaataccg catcaggcgc cattcgccat tcaggctgcg caactgttgg gaagggcgat
     9481 cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat
     9541 taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtgcca
     9601 agctg
//
