LOCUS       V032340                 9961 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V032340
VERSION     V032340
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     LTR             1..634
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     misc_feature    681..806
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
                     /label="HIV-1 Ψ"
     misc_feature    1303..1536
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
                     /label="RRE"
     CDS             1721..1765
                     /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    2028..2143
                     /note="central polypurine tract and central termination
                     sequence of HIV-1 (lacking the first T)"
                     /label="cPPT/CTS"
     enhancer        2201..2504
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        2505..2708
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             2826..4010
                     /label="CREB3L4(NM_130898)"
                     /note="CREB3L4(NM_130898)"
                     /gene="CREB3L4"
     CDS             4059..4769
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /transl_table=1
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK*"
                     /label="mCherry"
     promoter        4796..5295
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5316..5915
                     /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    5929..6517
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6724..7357
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7486..7502)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7510..7526
                     /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(7534..7564)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7579..7600
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7888..8476)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8647..9507)
                     /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(9508..9612)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9660..9794
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
ORIGIN
        1 tggaagggct aattcactcc caaagaagac aagatatcct tgatctgtgg atctaccaca
       61 cacaaggcta cttccctgat tagcagaact acacaccagg gccaggggtc agatatccac
      121 tgacctttgg atggtgctac aagctagtac cagttgagcc agataaggta gaagaggcca
      181 ataaaggaga gaacaccagc ttgttacacc ctgtgagcct gcatgggatg gatgacccgg
      241 agagagaagt gttagagtgg aggtttgaca gccgcctagc atttcatcac gtggcccgag
      301 agctgcatcc ggagtacttc aagaactgct gatatcgagc ttgctacaag ggactttccg
      361 ctggggactt tccagggagg cgtggcctgg gcgggactgg ggagtggcga gccctcagat
      421 cctgcatata agcagctgct ttttgcctgt actgggtctc tctggttaga ccagatctga
      481 gcctgggagc tctctggcta actagggaac ccactgctta agcctcaata aagcttgcct
      541 tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact ctggtaacta gagatccctc
      601 agaccctttt agtcagtgtg gaaaatctct agcagtggcg cccgaacagg gacttgaaag
      661 cgaaagggaa accagaggag ctctctcgac gcaggactcg gcttgctgaa gcgcgcacgg
      721 caagaggcga ggggcggcga ctggtgagta cgccaaaaat tttgactagc ggaggctaga
      781 aggagagaga tgggtgcgag agcgtcagta ttaagcgggg gagaattaga tcgcgatggg
      841 aaaaaattcg gttaaggcca gggggaaaga aaaaatataa attaaaacat atagtatggg
      901 caagcaggga gctagaacga ttcgcagtta atcctggcct gttagaaaca tcagaaggct
      961 gtagacaaat actgggacag ctacaaccat cccttcagac aggatcagaa gaacttagat
     1021 cattatataa tacagtagca accctctatt gtgtgcatca aaggatagag ataaaagaca
     1081 ccaaggaagc tttagacaag atagaggaag agcaaaacaa aagtaagacc accgcacagc
     1141 aagcggccgg ccgctgatct tcagacctgg aggaggagat atgagggaca attggagaag
     1201 tgaattatat aaatataaag tagtaaaaat tgaaccatta ggagtagcac ccaccaaggc
     1261 aaagagaaga gtggtgcaga gagaaaaaag agcagtggga ataggagctt tgttccttgg
     1321 gttcttggga gcagcaggaa gcactatggg cgcagcgtca atgacgctga cggtacaggc
     1381 cagacaatta ttgtctggta tagtgcagca gcagaacaat ttgctgaggg ctattgaggc
     1441 gcaacagcat ctgttgcaac tcacagtctg gggcatcaag cagctccagg caagaatcct
     1501 ggctgtggaa agatacctaa aggatcaaca gctcctgggg atttggggtt gctctggaaa
     1561 actcatttgc accactgctg tgccttggaa tgctagttgg agtaataaat ctctggaaca
     1621 gatttggaat cacacgacct ggatggagtg ggacagagaa attaacaatt acacaagctt
     1681 aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa aagaatgaac aagaattatt
     1741 ggaattagat aaatgggcaa gtttgtggaa ttggtttaac ataacaaatt ggctgtggta
     1801 tataaaatta ttcataatga tagtaggagg cttggtaggt ttaagaatag tttttgctgt
     1861 actttctata gtgaatagag ttaggcaggg atattcacca ttatcgtttc agacccacct
     1921 cccaaccccg aggggacccg acaggcccga aggaatagaa gaagaaggtg gagagagaga
     1981 cagagacaga tccattcgat tagtgaacgg atctcgacgg tatcgccttt aaaagaaaag
     2041 gggggattgg ggggtacagt gcaggggaaa gaatagtaga cataatagca acagacatac
     2101 aaactaaaga attacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgataagctt gggagttccg cgttacataa cttacggtaa
     2221 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     2281 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     2341 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     2401 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     2461 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     2521 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     2581 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     2641 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     2701 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     2761 tccatagaag acaccgactc tactagagga tcgctagcgc taccggactc agatctcgag
     2821 ccaccatgga tctcggaatc cctgacctgc tggacgcgtg gctggagccc ccagaggata
     2881 tcttctcgac aggatccgtc ctggagctgg gactccactg cccccctcca gaggttccgg
     2941 taactaggct acaggaacag ggactgcaag gctggaagtc cggtggggac cgtggctgtg
     3001 gccttcaaga gagtgagcct gaagatttct tgaagctttt cattgatccc aatgaggtgt
     3061 actgctcaga agcatctcct ggcagtgaca gtggcatctc tgaggacccc tgccatccag
     3121 acagtccccc tgcccccagg gcaaccagtt ctcctatgct ctatgaggtt gtctatgagg
     3181 caggggccct ggagaggatg cagggggaaa ctgggccaaa tgtaggcctt atctccatcc
     3241 agctagatca gtggagccca gcatttatgg tgcctgattc ctgcatggtc agtgagctgc
     3301 cctttgatgc tcatgcccac atcctgccca gagcaggcac cgtagcccca gtgccctgta
     3361 caaccctgct gccctgtcaa accctgttcc tgaccgatga ggagaagcgt ctgctggggc
     3421 aggaaggggt ttccctgccc tctcacctgc ccctcaccaa ggcagaggag agggtcctca
     3481 agaaggtcag gaggaaaatc cgtaacaagc agtcagctca ggacagtcgg cggcggaaga
     3541 aggagtacat tgatgggctg gagagcaggg tggcagcctg ttctgcacag aaccaagaat
     3601 tacagaaaaa agtccaggag ctggagaggc acaacatctc cttggtagct cagctccgcc
     3661 agctgcagac gctaattgct caaacttcca acaaagctgc ccagaccagc acttgtgttt
     3721 tgattcttct tttttccctg gctctcatca tcctgcccag cttcagtcca ttccagagtc
     3781 gaccagaagc tgggtctgag gattaccagc ctcacggagt gacttccaga aatatcctga
     3841 cccacaagga cgtaacagaa aatctggaga cccaagtggt agagtccaga ctgagggagc
     3901 cacctggagc caaggatgca aatggctcaa caaggacact gcttgagaag atgggaggga
     3961 agccaagacc cagtgggcgc atccggtccg tgctgcatgc agatgagatg gaattcggag
     4021 gtggcggtag tggtggcgga ggatccacgg tcgccaccat ggtgagcaag ggcgaggagg
     4081 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     4141 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     4201 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     4261 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     4321 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     4381 gcgtggtgac cgtgacccag gactcctccc tgcaggacgg cgagttcatc tacaaggtga
     4441 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     4501 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     4561 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     4621 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     4681 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     4741 ccggcggcat ggacgagctg tacaagtagc ggccgcgact ctagataatt ctaccgggta
     4801 ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc gctgggcact
     4861 tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg taggcgccaa
     4921 ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct agtcaggaag
     4981 ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc
     5041 actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg cctttggggc
     5101 agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg aaggggtggg
     5161 tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga
     5221 ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca
     5281 tctccgggcc tttcgacctg cagcccaagc ttaccatgac cgagtacaag cccacggtgc
     5341 gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg
     5401 actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc
     5461 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     5521 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     5581 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     5641 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5701 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5761 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5821 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5881 gcacctggtg catgacccgc aagcccggtg cctgaccgcg tctggaacaa tcaacctctg
     5941 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     6001 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     6061 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6121 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     6181 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     6241 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6301 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     6361 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6421 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6481 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     6541 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     6601 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     6661 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg
     6721 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     6781 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     6841 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     6901 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     6961 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     7021 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     7081 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     7141 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     7201 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7261 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7321 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     7381 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc
     7441 tagcgtttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7501 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7561 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7621 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7681 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7741 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7801 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7861 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7921 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     7981 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     8041 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     8101 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     8161 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     8221 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8281 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     8341 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8401 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8461 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8521 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8581 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8641 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8701 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8761 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8821 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8881 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8941 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     9001 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9061 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9121 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     9181 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9241 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9301 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9361 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9421 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9481 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9541 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9601 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca
     9661 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     9721 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9781 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     9841 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     9901 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
     9961 t
//
