LOCUS       V025919                10225 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025919
VERSION     V025919
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             2843..4240
                     /label="VIM(NM_003380)"
                     /note="VIM(NM_003380)"
                     /gene="VIM"
     CDS             4277..4996
                     /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"
     promoter        5013..5512
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5533..6132
                     /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    6146..6734
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6941..7574
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7703..7719)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7727..7743
                     /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(7751..7781)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7796..7817
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8105..8693)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8864..9724)
                     /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(9725..9829)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9877..10011
                     /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 ctcaagcttc gaattcgcca ccatgtccac caggtccgtg tcctcgtcct cctaccgcag
     2881 gatgttcggc ggcccgggca ccgcgagccg gccgagctcc agccggagct acgtgactac
     2941 gtccacccgc acctacagcc tgggcagcgc gctgcgcccc agcaccagcc gcagcctcta
     3001 cgcctcgtcc ccgggcggcg tgtatgccac gcgctcctct gccgtgcgcc tgcggagcag
     3061 cgtgcccggg gtgcggctcc tgcaggactc ggtggacttc tcgctggccg acgccatcaa
     3121 caccgagttc aagaacaccc gcaccaacga gaaggtggag ctgcaggagc tgaatgaccg
     3181 cttcgccaac tacatcgaca aggtgcgctt cctggagcag cagaataaga tcctgctggc
     3241 cgagctcgag cagctcaagg gccaaggcaa gtcgcgcctg ggggacctct acgaggagga
     3301 gatgcgggag ctgcgccggc aggtggacca gctaaccaac gacaaagccc gcgtcgaggt
     3361 ggagcgcgac aacctggccg aggacatcat gcgcctccgg gagaaattgc aggaggagat
     3421 gcttcagaga gaggaagccg aaaacaccct gcaatctttc agacaggatg ttgacaatgc
     3481 gtctctggca cgtcttgacc ttgaacgcaa agtggaatct ttgcaagaag agattgcctt
     3541 tttgaagaaa ctccacgaag aggaaatcca ggagctgcag gctcagattc aggaacagca
     3601 tgtccaaatc gatgtggatg tttccaagcc tgacctcacg gctgccctgc gtgacgtacg
     3661 tcagcaatat gaaagtgtgg ctgccaagaa cctgcaggag gcagaagaat ggtacaaatc
     3721 caagtttgct gacctctctg aggctgccaa ccggaacaat gacgccctgc gccaggcaaa
     3781 gcaggagtcc actgagtacc ggagacaggt gcagtccctc acctgtgaag tggatgccct
     3841 taaaggaacc aatgagtccc tggaacgcca gatgcgtgaa atggaagaga actttgccgt
     3901 tgaagctgct aactaccaag acactattgg ccgcctgcag gatgagattc agaatatgaa
     3961 ggaggaaatg gctcgtcacc ttcgtgaata ccaagacctg ctcaatgtta agatggccct
     4021 tgacattgag attgccacct acaggaagct gctggaaggc gaggagagca ggatttctct
     4081 gcctcttcca aacttttcct ccctgaacct gagggaaact aatctggatt cactccctct
     4141 ggttgatacc cactcaaaaa ggacacttct gattaagacg gttgaaacta gagatggaca
     4201 ggttatcaac gaaacttctc agcatcacga tgaccttgaa ggcggcggag ggagtggagg
     4261 agggggaagc ggatccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     4321 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     4381 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     4441 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     4501 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     4561 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     4621 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     4681 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     4741 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     4801 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     4861 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     4921 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     4981 cgagctgtac aagtagtcta gataattcta ccgggtaggg gaggcgcttt tcccaaggca
     5041 gtctggagca tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg
     5101 gcctcgcaca cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc
     5161 cttcgcgcca ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc
     5221 gtcgtgcagg acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag
     5281 caccgctgag caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc
     5341 cttcgctttc tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg
     5401 gctcaggggc ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca
     5461 aaagcgcacg tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag
     5521 cccaagctta ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     5581 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     5641 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     5701 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     5761 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     5821 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     5881 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     5941 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     6001 cccgccttcc tggagacctc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     6061 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     6121 cccggtgcct gaccgcgtct ggaacaatca acctctggat tacaaaattt gtgaaagatt
     6181 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     6241 tttgtatcat gctattgctt cccgtatggc tttcattttc tcctccttgt ataaatcctg
     6301 gttgctgtct ctttatgagg agttgtggcc cgttgtcagg caacgtggcg tggtgtgcac
     6361 tgtgtttgct gacgcaaccc ccactggttg gggcattgcc accacctgtc agctcctttc
     6421 cgggactttc gctttccccc tccctattgc cacggcggaa ctcatcgccg cctgccttgc
     6481 ccgctgctgg acaggggctc ggctgttggg cactgacaat tccgtggtgt tgtcggggaa
     6541 gctgacgtcc tttccatggc tgctcgcctg tgttgccacc tggattctgc gcgggacgtc
     6601 cttctgctac gtcccttcgg ccctcaatcc agcggacctt ccttcccgcg gcctgctgcc
     6661 ggctctgcgg cctcttccgc gtcttcgcct tcgccctcag acgagtcgga tctccctttg
     6721 ggccgcctcc ccgcctggaa ttaattctgc agtcgagacc tagaaaaaca tggagcaatc
     6781 acaagtagca atacagcagc taccaatgct gattgtgcct ggctagaagc acaagaggag
     6841 gaggaggtgg gttttccagt cacacctcag gtacctttaa gaccaatgac ttacaaggca
     6901 gctgtagatc ttagccactt tttaaaagaa aagaggggac tggaagggct aattcactcc
     6961 caacgaagac aagatatcct tgatctgtgg atctaccaca cacaaggcta cttccctgat
     7021 tagcagaact acacaccagg gccaggggtc agatatccac tgacctttgg atggtgctac
     7081 aagctagtac cagttgagcc agataaggta gaagaggcca ataaaggaga gaacaccagc
     7141 ttgttacacc ctgtgagcct gcatgggatg gatgacccgg agagagaagt gttagagtgg
     7201 aggtttgaca gccgcctagc atttcatcac gtggcccgag agctgcatcc ggagtacttc
     7261 aagaactgct gatatcgagc ttgctacaag ggactttccg ctggggactt tccagggagg
     7321 cgtggcctgg gcgggactgg ggagtggcga gccctcagat cctgcatata agcagctgct
     7381 ttttgcctgt actgggtctc tctggttaga ccagatctga gcctgggagc tctctggcta
     7441 actagggaac ccactgctta agcctcaata aagcttgcct tgagtgcttc aagtagtgtg
     7501 tgcccgtctg ttgtgtgact ctggtaacta gagatccctc agaccctttt agtcagtgtg
     7561 gaaaatctct agcagtagta gttcatgtca tcttattatt cagtatttat aacttgcaaa
     7621 gaaatgaata tcagagagtg agaggccttg acattgctag cgttttaccg tcgacctcta
     7681 gctagagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     7741 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt gcctaatgag
     7801 tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt
     7861 cgtgccagct gcattaatga atcggccaac gcgcggggag aggcggtttg cgtattgggc
     7921 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     7981 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     8041 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     8101 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     8161 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     8221 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     8281 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     8341 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     8401 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     8461 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     8521 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
     8581 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     8641 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     8701 ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt
     8761 tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa aaatgaagtt
     8821 ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     8881 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     8941 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     9001 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     9061 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     9121 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     9181 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     9241 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     9301 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     9361 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     9421 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     9481 tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     9541 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     9601 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     9661 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     9721 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     9781 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     9841 gaaaagtgcc acctgacgtc gacggatcgg gagatcaact tgtttattgc agcttataat
     9901 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     9961 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctggat caactggata
    10021 actcaagcta accaaaatca tcccaaactt cccaccccat accctattac cactgccaat
    10081 tacctgtggt ttcatttact ctaaacctgt gattcctctg aattattttc attttaaaga
    10141 aattgtattt gttaaatatg tactacaaac ttagtagttt ttaaagaaat tgtatttgtt
    10201 aaatatgtac tacaaactta gtagt
//
