LOCUS       V029868                10090 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V029868
VERSION     V029868
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             2815..4008
                     /label="B4GALT1(NM_001497)"
                     /note="B4GALT1(NM_001497)"
                     /gene="B4GALT1"
     CDS             4045..4764
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK*"
                     /label="EGFP"
     misc_feature    4793..5379
                     /note="internal ribosome entry site (IRES) of the
                     encephalomyocarditis virus (EMCV)"
                     /label="IRES2"
     CDS             5399..5998
                     /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    6012..6600
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6807..7440
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7568..7584)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7592..7608
                     /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(7616..7646)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7661..7682
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7970..8558)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8729..9589)
                     /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(9590..9694)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9742..9876
                     /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 tctatttccg gtgaattcgc caccatgagg
     2821 cttcgggagc cgctcctgag cggcagcgcc gcgatgccag gcgcgtccct acagcgggcc
     2881 tgccgcctgc tcgtggccgt ctgcgctctg caccttggcg tcaccctcgt ttactacctg
     2941 gctggccgcg acctgagccg cctgccccaa ctggtcggag tctccacacc gctgcagggc
     3001 ggctcgaaca gtgccgccgc catcgggcag tcctccgggg agctccggac cggaggggcc
     3061 cggccgccgc ctcctctagg cgcctcctcc cagccgcgcc cgggtggcga ctccagccca
     3121 gtcgtggatt ctggccctgg ccccgctagc aacttgacct cggtcccagt gccccacacc
     3181 accgcactgt cgctgcccgc ctgccctgag gagtccccgc tgcttgtggg ccccatgctg
     3241 attgagttta acatgcctgt ggacctggag ctcgtggcaa agcagaaccc aaatgtgaag
     3301 atgggcggcc gctatgcccc cagggactgc gtctctcctc acaaggtggc catcatcatt
     3361 ccattccgca accggcagga gcacctcaag tactggctat attatttgca cccagtcctg
     3421 cagcgccagc agctggacta tggcatctat gttatcaacc aggcgggaga cactatattc
     3481 aatcgtgcta agctcctcaa tgttggcttt caagaagcct tgaaggacta tgactacacc
     3541 tgctttgtgt ttagtgacgt ggacctcatt ccaatgaatg accataatgc gtacaggtgt
     3601 ttttcacagc cacggcacat ttccgttgca atggataagt ttggattcag cctaccttat
     3661 gttcagtatt ttggaggtgt ctctgctcta agtaaacaac agtttctaac catcaatgga
     3721 tttcctaata attattgggg ctggggagga gaagatgatg acatttttaa cagattagtt
     3781 tttagaggca tgtctatatc tcgcccaaat gctgtggtcg ggaggtgtcg catgatccgc
     3841 cactcaagag acaagaaaaa tgaacccaat cctcagaggt ttgaccgaat tgcacacaca
     3901 aaggagacaa tgctctctga tggtttgaac tcactcacct accaggtgct ggatgtacag
     3961 agatacccat tgtataccca aatcacagtg gacatcggga caccgagcct cgagggcggt
     4021 ggaggaagcg gcggaggtgg atctatggtg agcaagggcg aggagctgtt caccggggtg
     4081 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     4141 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     4201 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     4261 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     4321 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     4381 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     4441 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     4501 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     4561 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     4621 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     4681 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     4741 ggcatggacg agctgtacaa gtagactagt tctagagcgg ccgcggatcc cgcccctctc
     4801 cctccccccc ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg
     4861 tctatatgtt attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg
     4921 gccctgtctt cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag
     4981 gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt
     5041 ctgtagcgac cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc
     5101 aaaagccacg tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga
     5161 gttggatagt tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga
     5221 aggatgccca gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct
     5281 ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg
     5341 ttttcctttg aaaaacacga tgataagctt gccacaaccc acaaggagac gaccttccat
     5401 gaccgagtac aagcccacgg tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg
     5461 caccctcgcc gccgcgttcg ccgactaccc cgccacgcgc cacaccgtcg acccggaccg
     5521 ccacatcgag cgggtcaccg agctgcaaga actcttcctc acgcgcgtcg ggctcgacat
     5581 cggcaaggtg tgggtcgcgg acgacggcgc cgcggtggcg gtctggacca cgccggagag
     5641 cgtcgaagcg ggggcggtgt tcgccgagat cggcccgcgc atggccgagt tgagcggttc
     5701 ccggctggcc gcgcagcaac agatggaagg cctcctggcg ccgcaccggc ccaaggagcc
     5761 cgcgtggttc ctggccaccg tcggcgtctc gcccgaccac cagggcaagg gtctgggcag
     5821 cgccgtcgtg ctccccggag tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga
     5881 gacctccgcg ccccgcaacc tccccttcta cgagcggctc ggcttcaccg tcaccgccga
     5941 cgtcgaggtg cccgaaggac cgcgcacctg gtgcatgacc cgcaagcccg gtgcctagac
     6001 gcgtctggaa caatcaacct ctggattaca aaatttgtga aagattgact ggtattctta
     6061 actatgttgc tccttttacg ctatgtggat acgctgcttt aatgcctttg tatcatgcta
     6121 ttgcttcccg tatggctttc attttctcct ccttgtataa atcctggttg ctgtctcttt
     6181 atgaggagtt gtggcccgtt gtcaggcaac gtggcgtggt gtgcactgtg tttgctgacg
     6241 caacccccac tggttggggc attgccacca cctgtcagct cctttccggg actttcgctt
     6301 tccccctccc tattgccacg gcggaactca tcgccgcctg ccttgcccgc tgctggacag
     6361 gggctcggct gttgggcact gacaattccg tggtgttgtc ggggaagctg acgtcctttc
     6421 catggctgct cgcctgtgtt gccacctgga ttctgcgcgg gacgtccttc tgctacgtcc
     6481 cttcggccct caatccagcg gaccttcctt cccgcggcct gctgccggct ctgcggcctc
     6541 ttccgcgtct tcgccttcgc cctcagacga gtcggatctc cctttgggcc gcctccccgc
     6601 ctggaattaa ttctgcagtc gagacctaga aaaacatgga gcaatcacaa gtagcaatac
     6661 agcagctacc aatgctgatt gtgcctggct agaagcacaa gaggaggagg aggtgggttt
     6721 tccagtcaca cctcaggtac ctttaagacc aatgacttac aaggcagctg tagatcttag
     6781 ccacttttta aaagaaaaga ggggactgga agggctaatt cactcccaac gaagacaaga
     6841 tatccttgat ctgtggatct accacacaca aggctacttc cctgattagc agaactacac
     6901 accagggcca ggggtcagat atccactgac ctttggatgg tgctacaagc tagtaccagt
     6961 tgagccagat aaggtagaag aggccaataa aggagagaac accagcttgt tacaccctgt
     7021 gagcctgcat gggatggatg acccggagag agaagtgtta gagtggaggt ttgacagccg
     7081 cctagcattt catcacgtgg cccgagagct gcatccggag tacttcaaga actgctgata
     7141 tcgagcttgc tacaagggac tttccgctgg ggactttcca gggaggcgtg gcctgggcgg
     7201 gactggggag tggcgagccc tcagatcctg catataagca gctgcttttt gcctgtactg
     7261 ggtctctctg gttagaccag atctgagcct gggagctctc tggctaacta gggaacccac
     7321 tgcttaagcc tcaataaagc ttgccttgag tgcttcaagt agtgtgtgcc cgtctgttgt
     7381 gtgactctgg taactagaga tccctcagac ccttttagtc agtgtggaaa atctctagca
     7441 gtagtagttc atgtcatctt attattcagt atttataact tgcaaagaaa tgaatatcag
     7501 agagtgagag gccttgacat tgctagcgtt taccgtcgac ctctagctag agcttggcgt
     7561 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     7621 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     7681 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     7741 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     7801 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     7861 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     7921 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     7981 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     8041 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     8101 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     8161 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     8221 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     8281 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     8341 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     8401 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     8461 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     8521 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     8581 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     8641 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     8701 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     8761 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     8821 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     8881 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     8941 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     9001 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     9061 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     9121 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     9181 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     9241 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     9301 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     9361 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     9421 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     9481 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     9541 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     9601 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     9661 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     9721 acgtcgacgg atcgggagat caacttgttt attgcagctt ataatggtta caaataaagc
     9781 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     9841 tccaaactca tcaatgtatc ttatcatgtc tggatcaact ggataactca agctaaccaa
     9901 aatcatccca aacttcccac cccataccct attaccactg ccaattacct gtggtttcat
     9961 ttactctaaa cctgtgattc ctctgaatta ttttcatttt aaagaaattg tatttgttaa
    10021 atatgtacta caaacttagt agtttttaaa gaaattgtat ttgttaaata tgtactacaa
    10081 acttagtagt
//
