LOCUS       V025329                10063 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V025329
VERSION     V025329
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..3559
                     /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"
     CDS             3629..4831
                     /label="En1(NM_010133)"
                     /note="En1(NM_010133)"
                     /gene="En1"
     promoter        4851..5350
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5371..5970
                     /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    5984..6572
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6779..7412
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7541..7557)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7565..7581
                     /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(7589..7619)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7634..7655
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7943..8531)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8702..9562)
                     /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(9563..9667)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9715..9849
                     /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 ccatggtgag caagggcgag gagctgttca ccggggtggt
     2881 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2941 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     3001 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     3061 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     3121 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     3181 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     3241 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     3301 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     3361 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     3421 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     3481 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3541 catggacgag ctgtacaagt ccggactcag atctcgaggc ggcggaggga gtggaggagg
     3601 gggaagcgga ggcggaggga gtggatccat ggaagaacag cagccggagc ctaaaagtca
     3661 gcgcgactcg ggcctcggcg cggtggcagc ggcggccccg agcggcctca gtctgagtct
     3721 gagcccagga gccagcggca gcagcggcag cgatggagac agcgtgccgg tgtccccgca
     3781 gccagcgccc ccgtcgcctc ctgcggcacc ctgtctgccg cccctggccc atcacccgca
     3841 cctccccccg catcccccgc ccccgccgcc gccgccgccg ccgccaccgc agcatctcgc
     3901 ggcgcctgct caccagccgc agcccgcggc ccagctgcac cgcaccacca actttttcat
     3961 cgataacatc ctaaggcccg atttcggttg caaaaaggaa cagcccctgc ctcagctcct
     4021 ggtggcttcg gctgcagccg gaggaggcgc agcagcagga ggaggaagcc gcgtggagcg
     4081 tgaccgaggc cagactggtg caggtagaga ccccgttcac tctctgggca cacgagcttc
     4141 gggggctgcc tcgctcttgt gtgctccaga tgcgaactgt ggcccacccg acggctccca
     4201 gcccgccacc gctgtcagcg ccggcgcatc caaagccggg aacccggctg ctgcggcggc
     4261 cgcggccgca gcagcggctg cagcggcagt ggcggcagcg gcggcagcag cctcgaagcc
     4321 ctcggacagt ggcggtggta gtggaggcaa cgcggggagt cccggggcgc agggcgccaa
     4381 gttcccggaa cacaaccctg cgatcctact catgggttcg gctaacggtg ggccggtggt
     4441 caagactgac tcacagcaac ccctagtgtg gcccgcctgg gtctactgca cacgctattc
     4501 ggaccgtccg tcctctggtc cacgcaccag gaagctaaag aagaaaaaga acgagaagga
     4561 agacaagcgg ccgcggacgg cgttcacggc cgagcagctg cagagactca aggcggagtt
     4621 ccaggcaaac cgctatatca cggagcagcg gcgacagacc ctcgcccagg agctcagcct
     4681 gaatgagtcc cagatcaaga tctggttcca aaacaagcgt gccaagatca agaaagccac
     4741 cggcatcaag aacggcctgg cgctgcacct catggcccag ggactgtaca accactctac
     4801 caccacggtt caggacaaag acgagagcga gtagtctaga taattctacc gggtagggga
     4861 ggcgcttttc ccaaggcagt ctggagcatg cgctttagca gccccgctgg gcacttggcg
     4921 ctacacaagt ggcctctggc ctcgcacaca ttccacatcc accggtaggc gccaaccggc
     4981 tccgttcttt ggtggcccct tcgcgccacc ttctactcct cccctagtca ggaagttccc
     5041 ccccgccccg cagctcgcgt cgtgcaggac gtgacaaatg gaagtagcac gtctcactag
     5101 tctcgtgcag atggacagca ccgctgagca atggaagcgg gtaggccttt ggggcagcgg
     5161 ccaatagcag ctttgctcct tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg
     5221 gggcgggctc aggggcgggc tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc
     5281 ggcattctgc acgcttcaaa agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc
     5341 gggcctttcg acctgcagcc caagcttacc atgaccgagt acaagcccac ggtgcgcctc
     5401 gccacccgcg acgacgtccc cagggccgta cgcaccctcg ccgccgcgtt cgccgactac
     5461 cccgccacgc gccacaccgt cgatccggac cgccacatcg agcgggtcac cgagctgcaa
     5521 gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc
     5581 gccgcggtgg cggtctggac cacgccggag agcgtcgaag cgggggcggt gttcgccgag
     5641 atcggcccgc gcatggccga gttgagcggt tcccggctgg ccgcgcagca acagatggaa
     5701 ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc
     5761 tcgcccgacc accagggcaa gggtctgggc agcgccgtcg tgctccccgg agtggaggcg
     5821 gccgagcgcg ccggggtgcc cgccttcctg gagacctccg cgccccgcaa cctccccttc
     5881 tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc
     5941 tggtgcatga cccgcaagcc cggtgcctga ccgcgtctgg aacaatcaac ctctggatta
     6001 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     6061 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     6121 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     6181 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     6241 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     6301 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     6361 cgtggtgttg tcggggaagc tgacgtcctt tccatggctg ctcgcctgtg ttgccacctg
     6421 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     6481 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     6541 gagtcggatc tccctttggg ccgcctcccc gcctggaatt aattctgcag tcgagaccta
     6601 gaaaaacatg gagcaatcac aagtagcaat acagcagcta ccaatgctga ttgtgcctgg
     6661 ctagaagcac aagaggagga ggaggtgggt tttccagtca cacctcaggt acctttaaga
     6721 ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa gaggggactg
     6781 gaagggctaa ttcactccca acgaagacaa gatatccttg atctgtggat ctaccacaca
     6841 caaggctact tccctgatta gcagaactac acaccagggc caggggtcag atatccactg
     6901 acctttggat ggtgctacaa gctagtacca gttgagccag ataaggtaga agaggccaat
     6961 aaaggagaga acaccagctt gttacaccct gtgagcctgc atgggatgga tgacccggag
     7021 agagaagtgt tagagtggag gtttgacagc cgcctagcat ttcatcacgt ggcccgagag
     7081 ctgcatccgg agtacttcaa gaactgctga tatcgagctt gctacaaggg actttccgct
     7141 ggggactttc cagggaggcg tggcctgggc gggactgggg agtggcgagc cctcagatcc
     7201 tgcatataag cagctgcttt ttgcctgtac tgggtctctc tggttagacc agatctgagc
     7261 ctgggagctc tctggctaac tagggaaccc actgcttaag cctcaataaa gcttgccttg
     7321 agtgcttcaa gtagtgtgtg cccgtctgtt gtgtgactct ggtaactaga gatccctcag
     7381 acccttttag tcagtgtgga aaatctctag cagtagtagt tcatgtcatc ttattattca
     7441 gtatttataa cttgcaaaga aatgaatatc agagagtgag aggccttgac attgctagcg
     7501 ttttaccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     7561 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     7621 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     7681 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     7741 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     7801 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     7861 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     7921 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     7981 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     8041 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     8101 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     8161 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     8221 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     8281 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     8341 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     8401 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     8461 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     8521 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     8581 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     8641 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     8701 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     8761 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     8821 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     8881 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     8941 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     9001 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     9061 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     9121 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     9181 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     9241 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     9301 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     9361 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     9421 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     9481 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     9541 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     9601 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     9661 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatcaacttg
     9721 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     9781 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     9841 gtctggatca actggataac tcaagctaac caaaatcatc ccaaacttcc caccccatac
     9901 cctattacca ctgccaatta cctgtggttt catttactct aaacctgtga ttcctctgaa
     9961 ttattttcat tttaaagaaa ttgtatttgt taaatatgta ctacaaactt agtagttttt
    10021 aaagaaattg tatttgttaa atatgtacta caaacttagt agt
//
