LOCUS       V031798                10006 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031798
VERSION     V031798
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             2807..3523
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="Aequorea coerulescens GFP"
                     /transl_table=1
                     /translation="MVSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDD
                     GNYKSRAEVKFEGDTLVNRIELTGTDFKEDGNILGNKMEYNYNAHNVYIMTDKAKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYF
                     GFVTAAAITHGMDELYK"
                     /label="AcGFP1"
     CDS             3572..4768
                     /label="B4galt1(NM_022305)"
                     /note="B4galt1(NM_022305)"
                     /gene="B4galt1"
     promoter        4841..5340
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5361..5960
                     /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    5974..6562
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6769..7402
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7531..7547)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7555..7571
                     /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(7579..7609)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7624..7645
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7933..8521)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8692..9552)
                     /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(9553..9657)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9705..9839
                     /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 tctaccggtc gccaccatgg tgagcaaggg
     2821 cgccgagctg ttcaccggca tcgtgcccat cctgatcgag ctgaatggcg atgtgaatgg
     2881 ccacaagttc agcgtgagcg gcgagggcga gggcgatgcc acctacggca agctgaccct
     2941 gaagttcatc tgcaccaccg gcaagctgcc tgtgccctgg cccaccctgg tgaccaccct
     3001 gagctacggc gtgcagtgct tctcacgcta ccccgatcac atgaagcagc acgacttctt
     3061 caagagcgcc atgcctgagg gctacatcca ggagcgcacc atcttcttcg aggatgacgg
     3121 caactacaag tcgcgcgccg aggtgaagtt cgagggcgat accctggtga atcgcatcga
     3181 gctgaccggc accgatttca aggaggatgg caacatcctg ggcaataaga tggagtacaa
     3241 ctacaacgcc cacaatgtgt acatcatgac cgacaaggcc aagaatggca tcaaggtgaa
     3301 cttcaagatc cgccacaaca tcgaggatgg cagcgtgcag ctggccgacc actaccagca
     3361 gaataccccc atcggcgatg gccctgtgct gctgcccgat aaccactacc tgtccaccca
     3421 gagcgccctg tccaaggacc ccaacgagaa gcgcgatcac atgatctact tcggcttcgt
     3481 gaccgccgcc gccatcaccc acggcatgga tgagctgtac aagtccggac tcagaggagg
     3541 tggaggtagt ggaggaggcg gaagcctcga gatgaggttt cgtgagcagt tcctgggcgg
     3601 cagcgccgcg atgccgggcg cgaccctgca gcgggcctgc cgcctgctcg tggccgtctg
     3661 cgcgctgcac ctcggcgtca ccctcgtcta ttacctctct ggccgcgatc tgagccgcct
     3721 gccccagttg gtcggagtct cctctacact gcagggcggc acgaacggcg ccgcagccag
     3781 caagcagccc ccaggagagc agcggccgcg gggtgcgcgg ccgccgcctc ctttaggcgt
     3841 ctccccgaag cctcgcccgg gtctcgactc cagccctggt gcagcttctg gccccggctt
     3901 gaagagcaac ttgtcttcgt tgccagtgcc caccaccact ggactgttgt cgctgccagc
     3961 ttgccctgag gagtccccgc tgctcgttgg ccccatgctg attgacttta atattgctgt
     4021 ggatctggag cttttggcaa agaagaaccc agagataaag acgggcggcc gttactcccc
     4081 caaggactgt gtctctcctc acaaggtggc catcatcatc ccattccgta accggcagga
     4141 gcatctcaaa tactggctgt attatttgca tcccatcctt cagcgccagc aactcgacta
     4201 tggcatctac gtcatcaatc aggctggaga caccatgttc aatcgagcta agctgctcaa
     4261 tattggcttt caagaggcct tgaaggacta tgattacaac tgctttgtgt tcagtgatgt
     4321 ggacctcatt ccgatggacg accgtaatgc ctacaggtgt ttttcgcagc cacggcacat
     4381 ttctgttgca atggacaagt tcgggtttag cctgccatat gttcagtatt ttggaggtgt
     4441 ctctgctctc agtaaacaac agtttcttgc catcaatgga ttccctaata attattgggg
     4501 ttggggagga gaagatgacg acatttttaa cagattagtt cataaaggca tgtctatatc
     4561 acgtccaaat gctgtagtag ggaggtgtcg aatgatccgg cattcaagag acaagaaaaa
     4621 tgagcccaat cctcagaggt ttgaccggat cgcacataca aaggaaacga tgcgcttcga
     4681 tggtttgaac tcacttacct acaaggtgtt ggatgtacag agatacccgt tatataccca
     4741 aatcacagtg gacatcggga caccgagata ggaattctgc agtcgacggt accgcgggcc
     4801 cgggatccac cggatctaga taactgatca taattctacc gggtagggga ggcgcttttc
     4861 ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt
     4921 ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt
     4981 ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg
     5041 cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag
     5101 atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag
     5161 ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc
     5221 aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc
     5281 acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg
     5341 acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg
     5401 acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc
     5461 gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc
     5521 tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg
     5581 cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc
     5641 gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg
     5701 cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc
     5761 accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg
     5821 ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc
     5881 tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga
     5941 cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac ctctggatta caaaatttgt
     6001 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     6061 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     6121 aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg
     6181 gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag
     6241 ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc
     6301 tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg
     6361 tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg gattctgcgc
     6421 gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc
     6481 ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc
     6541 tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta gaaaaacatg
     6601 gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg ctagaagcac
     6661 aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga ccaatgactt
     6721 acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg gaagggctaa
     6781 ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca caaggctact
     6841 tccctgatta gcagaactac acaccagggc caggggtcag atatccactg acctttggat
     6901 ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat aaaggagaga
     6961 acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag agagaagtgt
     7021 tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag ctgcatccgg
     7081 agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct ggggactttc
     7141 cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc tgcatataag
     7201 cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc ctgggagctc
     7261 tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg agtgcttcaa
     7321 gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag acccttttag
     7381 tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca gtatttataa
     7441 cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg ttttaccgtc
     7501 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     7561 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     7621 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     7681 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     7741 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     7801 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     7861 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     7921 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     7981 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     8041 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     8101 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     8161 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     8221 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     8281 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     8341 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     8401 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     8461 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     8521 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     8581 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     8641 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     8701 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     8761 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     8821 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     8881 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     8941 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     9001 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     9061 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     9121 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     9181 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     9241 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     9301 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     9361 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     9421 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     9481 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     9541 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     9601 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     9661 atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg tttattgcag
     9721 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     9781 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     9841 actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac cctattacca
     9901 ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa ttattttcat
     9961 tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagt
//
