LOCUS       V044785                10168 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044785
VERSION     V044785
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             2846..2863
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /transl_table=1
                     /translation="HHHHHH"
                     /label="6xHis"
     CDS             2864..4924
                     /label="FLT1(NM_001159920)"
                     /note="FLT1(NM_001159920)"
                     /gene="FLT1"
     promoter        4956..5455
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5476..6075
                     /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    6089..6677
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             6884..7517
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7646..7662)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7670..7686
                     /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(7694..7724)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7739..7760
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8048..8636)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8807..9667)
                     /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(9668..9772)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    9820..9954
                     /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 ccatgcatca ccatcaccat cacatggtca gctactggga
     2881 caccggggtc ctgctgtgcg cgctgctcag ctgtctgctt ctcacaggat ctagttcagg
     2941 ttcaaaatta aaagatcctg aactgagttt aaaaggcacc cagcacatca tgcaagcagg
     3001 ccagacactg catctccaat gcagggggga agcagcccat aaatggtctt tgcctgaaat
     3061 ggtgagtaag gaaagcgaaa ggctgagcat aactaaatct gcctgtggaa gaaatggcaa
     3121 acaattctgc agtactttaa ccttgaacac agctcaagca aaccacactg gcttctacag
     3181 ctgcaaatat ctagctgtac ctacttcaaa gaagaaggaa acagaatctg caatctatat
     3241 atttattagt gatacaggta gacctttcgt agagatgtac agtgaaatcc ccgaaattat
     3301 acacatgact gaaggaaggg agctcgtcat tccctgccgg gttacgtcac ctaacatcac
     3361 tgttacttta aaaaagtttc cacttgacac tttgatccct gatggaaaac gcataatctg
     3421 ggacagtaga aagggcttca tcatatcaaa tgcaacgtac aaagaaatag ggcttctgac
     3481 ctgtgaagca acagtcaatg ggcatttgta taagacaaac tatctcacac atcgacaaac
     3541 caatacaatc atagatgtcc aaataagcac accacgccca gtcaaattac ttagaggcca
     3601 tactcttgtc ctcaattgta ctgctaccac tcccttgaac acgagagttc aaatgacctg
     3661 gagttaccct gatgaaaaaa ataagagagc ttccgtaagg cgacgaattg accaaagcaa
     3721 ttcccatgcc aacatattct acagtgttct tactattgac aaaatgcaga acaaagacaa
     3781 aggactttat acttgtcgtg taaggagtgg accatcattc aaatctgtta acacctcagt
     3841 gcatatatat gataaagcat tcatcactgt gaaacatcga aaacagcagg tgcttgaaac
     3901 cgtagctggc aagcggtctt accggctctc tatgaaagtg aaggcatttc cctcgccgga
     3961 agttgtatgg ttaaaagatg ggttacctgc gactgagaaa tctgctcgct atttgactcg
     4021 tggctactcg ttaattatca aggacgtaac tgaagaggat gcagggaatt atacaatctt
     4081 gctgagcata aaacagtcaa atgtgtttaa aaacctcact gccactctaa ttgtcaatgt
     4141 gaaaccccag atttacgaaa aggccgtgtc atcgtttcca gacccggctc tctacccact
     4201 gggcagcaga caaatcctga cttgtaccgc atatggtatc cctcaaccta caatcaagtg
     4261 gttctggcac ccctgtaacc ataatcattc cgaagcaagg tgtgactttt gttccaataa
     4321 tgaagagtcc tttatcctgg atgctgacag caacatggga aacagaattg agagcatcac
     4381 tcagcgcatg gcaataatag aaggaaagaa taagatggct agcaccttgg ttgtggctga
     4441 ctctagaatt tctggaatct acatttgcat agcttccaat aaagttggga ctgtgggaag
     4501 aaacataagc ttttatatca cagatgtgcc aaatgggttt catgttaact tggaaaaaat
     4561 gccgacggaa ggagaggacc tgaaactgtc ttgcacagtt aacaagttct tatacagaga
     4621 cgttacttgg attttactgc ggacagttaa taacagaaca atgcactaca gtattagcaa
     4681 gcaaaaaatg gccatcacta aggagcactc catcactctt aatcttacca tcatgaatgt
     4741 ttccctgcaa gattcaggca cctatgcctg cagagccagg aatgtataca caggggaaga
     4801 aatcctccag aagaaagaaa ttacaatcag aggtgagcac tgcaacaaaa aggctgtttt
     4861 ctctcggatc tccaaattta aaagcacaag gaatgattgt accacacaaa gtaatgtaaa
     4921 acattaagga tcccgcgact ctagataatt ctaccgggta ggggaggcgc ttttcccaag
     4981 gcagtctgga gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct
     5041 ctggcctcgc acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg
     5101 ccccttcgcg ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct
     5161 cgcgtcgtgc aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga
     5221 cagcaccgct gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg
     5281 ctccttcgct ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg
     5341 cgggctcagg ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct
     5401 tcaaaagcgc acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg
     5461 cagcccaagc ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac
     5521 gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac
     5581 accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg
     5641 cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc
     5701 tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg
     5761 gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg
     5821 caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag
     5881 ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg
     5941 gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc
     6001 ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc
     6061 aagcccggtg cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag
     6121 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     6181 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6241 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6301 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6361 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6421 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6481 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6541 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6601 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6661 ttgggccgcc tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca
     6721 atcacaagta gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag
     6781 gaggaggagg tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag
     6841 gcagctgtag atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac
     6901 tcccaacgaa gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct
     6961 gattagcaga actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc
     7021 tacaagctag taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc
     7081 agcttgttac accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag
     7141 tggaggtttg acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac
     7201 ttcaagaact gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg
     7261 aggcgtggcc tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct
     7321 gctttttgcc tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg
     7381 ctaactaggg aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt
     7441 gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt
     7501 gtggaaaatc tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc
     7561 aaagaaatga atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct
     7621 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     7681 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     7741 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     7801 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     7861 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     7921 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     7981 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     8041 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     8101 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     8161 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     8221 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     8281 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     8341 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     8401 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     8461 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     8521 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     8581 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     8641 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     8701 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     8761 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     8821 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     8881 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     8941 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     9001 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     9061 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     9121 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     9181 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     9241 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     9301 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     9361 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     9421 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     9481 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     9541 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     9601 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     9661 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     9721 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     9781 cccgaaaagt gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat
     9841 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     9901 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg
     9961 ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc
    10021 aattacctgt ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa
    10081 agaaattgta tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt
    10141 gttaaatatg tactacaaac ttagtagt
//
