LOCUS       V031751                10019 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031751
VERSION     V031751
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..4028
                     /label="PHLDA1(NM_007350)"
                     /note="PHLDA1(NM_007350)"
                     /gene="PHLDA1"
     CDS             4071..4790
                     /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        4807..5306
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5327..5926
                     /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    5940..6528
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6735..7368
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7497..7513)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7521..7537
                     /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(7545..7575)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7590..7611
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7899..8487)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8658..9518)
                     /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(9519..9623)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9671..9805
                     /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 ccaccatgag gcgtgcgccg gctgccgagc gcctcttgga gctgggcttt cccccgcggt
     2881 gcgggcgcca ggagccgcct tttccgctgg gtgtcactcg ggggtgggga agatggccca
     2941 ttcaaaagcg ccgcgagggg gcccggccag tgcccttcag tgagcgctcg caagaggacg
     3001 gcagaggccc ggcagctcgg agctccggga ccttgtggcg catcaggacg cggctgtccc
     3061 tctgccggga cccagagccg ccgccgccgc tctgcctcct gcgtgttagc ctcctctgcg
     3121 cgctccgggc aggcggccgt gggagccgct ggggcgagga cggcgcgagg ctgctgctgc
     3181 tgcccccggc ccgcgcggct ggaaacggag aggccgagcc aagcggcggc ccctcttatg
     3241 ctgggaggat gctggagagt agcggctgca aagcgctgaa ggagggcgtg ctggagaagc
     3301 gcagcgacgg gttgttgcag ctctggaaga aaaagtgttg catcctcacc gaggaagggc
     3361 tgctgcttat cccgcccaag cagctgcaac accagcagca gcagcagcaa cagcagcagc
     3421 agcagcagca acaacagccc gggcaggggc cggccgagcc gtcccaaccc agtggccccg
     3481 ctgtcgccag cctcgagccg ccggtcaagc tcaaggaact gcacttctcc aacatgaaga
     3541 ccgtggactg tgtggagcgc aagggcaagt acatgtactt cactgtggtg atggcagagg
     3601 gcaaggagat cgactttcgg tgcccgcaag accagggctg gaacgccgag atcacgctgc
     3661 agatggtgca gtacaagaat cgtcaggcca tcctggcggt caaatccacg cggcagaagc
     3721 agcagcacct ggtccagcag cagcccccct cgcagccgca gccgcagccg cagctccagc
     3781 cccaacccca gcctcagcct cagccgcaac cccagcccca atcacaaccc cagcctcagc
     3841 cccaacccaa gcctcagccc cagcagctcc acccgtatcc gcatccacat ccacatccac
     3901 actctcatcc tcactcgcac ccacaccctc acccgcaccc gcatccgcac caaataccgc
     3961 acccacaccc acagccgcac tcgcagccgc acgggcaccg gcttctccgc agcacctcca
     4021 actctgccgg atccggcggc ggagggagtg gaggaggggg aagcgccacc atggtgagca
     4081 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
     4141 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
     4201 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     4261 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
     4321 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     4381 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     4441 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     4501 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
     4561 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
     4621 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     4681 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     4741 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag tctagataat
     4801 tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct ttagcagccc
     4861 cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc acatccaccg
     4921 gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct actcctcccc
     4981 tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga caaatggaag
     5041 tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg aagcgggtag
     5101 gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct cagaggctgg
     5161 gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg ggcgcccgaa
     5221 ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc gcgctgttct
     5281 cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga ccgagtacaa
     5341 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     5401 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     5461 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     5521 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     5581 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     5641 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     5701 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     5761 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga cctccgcgcc
     5821 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     5881 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaccgc gtctggaaca
     5941 atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac tatgttgctc
     6001 cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt gcttcccgta
     6061 tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat gaggagttgt
     6121 ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca acccccactg
     6181 gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc cccctcccta
     6241 ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg gctcggctgt
     6301 tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca tggctgctcg
     6361 cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct tcggccctca
     6421 atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt ccgcgtcttc
     6481 gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct ggaattaatt
     6541 ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag cagctaccaa
     6601 tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc cagtcacacc
     6661 tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc actttttaaa
     6721 agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata tccttgatct
     6781 gtggatctac cacacacaag gctacttccc tgattagcag aactacacac cagggccagg
     6841 ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg agccagataa
     6901 ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga gcctgcatgg
     6961 gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc tagcatttca
     7021 tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc gagcttgcta
     7081 caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga ctggggagtg
     7141 gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg tctctctggt
     7201 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     7261 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     7321 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt agtagttcat
     7381 gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag agtgagaggc
     7441 cttgacattg ctagcgtttt accgtcgacc tctagctaga gcttggcgta atcatggtca
     7501 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     7561 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     7621 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     7681 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     7741 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     7801 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     7861 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     7921 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     7981 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     8041 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     8101 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     8161 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     8221 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     8281 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     8341 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     8401 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     8461 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     8521 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     8581 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     8641 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     8701 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     8761 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     8821 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     8881 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     8941 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     9001 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     9061 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     9121 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     9181 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     9241 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     9301 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     9361 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     9421 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     9481 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     9541 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     9601 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     9661 tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     9721 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     9781 caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa atcatcccaa
     9841 acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt tactctaaac
     9901 ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa tatgtactac
     9961 aaacttagta gtttttaaag aaattgtatt tgttaaatat gtactacaaa cttagtagt
//
