LOCUS       V020823                10458 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V020823
VERSION     V020823
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             2821..5184
                     /label="ITGB3(NM_000212)"
                     /note="ITGB3(NM_000212)"
                     /gene="ITGB3"
     CDS             5191..5214
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     promoter        5246..5745
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             5766..6365
                     /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    6379..6967
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             7174..7807
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(7936..7952)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7960..7976
                     /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(7984..8014)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    8029..8050
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(8338..8926)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(9097..9957)
                     /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(9958..10062)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    10110..10244
                     /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 atgcgagcgc ggccgcggcc ccggccgctc tgggcgactg tgctggcgct gggggcgctg
     2881 gcgggcgttg gcgtaggagg gcccaacatc tgtaccacgc gaggtgtgag ctcctgccag
     2941 cagtgcctgg ctgtgagccc catgtgtgcc tggtgctctg atgaggccct gcctctgggc
     3001 tcacctcgct gtgacctgaa ggagaatctg ctgaaggata actgtgcccc agaatccatc
     3061 gagttcccag tgagtgaggc ccgagtacta gaggacaggc ccctcagcga caagggctct
     3121 ggagacagct cccaggtcac tcaagtcagt ccccagagga ttgcactccg gctccggcca
     3181 gatgattcga agaatttctc catccaagtg cggcaggtgg aggattaccc tgtggacatc
     3241 tactacttga tggacctgtc ttactccatg aaggatgatc tgtggagcat ccagaacctg
     3301 ggtaccaagc tggccaccca gatgcgaaag ctcaccagta acctgcggat tggcttcggg
     3361 gcatttgtgg acaagcctgt gtcaccatac atgtatatct ccccaccaga ggccctcgaa
     3421 aacccctgct atgatatgaa gaccacctgc ttgcccatgt ttggctacaa acacgtgctg
     3481 acgctaactg accaggtgac ccgcttcaat gaggaagtga agaagcagag tgtgtcacgg
     3541 aaccgagatg ccccagaggg tggctttgat gccatcatgc aggctacagt ctgtgatgaa
     3601 aagattggct ggaggaatga tgcatcccac ttgctggtgt ttaccactga tgccaagact
     3661 catatagcat tggacggaag gctggcaggc attgtccagc ctaatgacgg gcagtgtcat
     3721 gttggtagtg acaatcatta ctctgcctcc actaccatgg attatccctc tttggggctg
     3781 atgactgaga agctatccca gaaaaacatc aatttgatct ttgcagtgac tgaaaatgta
     3841 gtcaatctct atcagaacta tagtgagctc atcccaggga ccacagttgg ggttctgtcc
     3901 atggattcca gcaatgtcct ccagctcatt gttgatgctt atgggaaaat ccgttctaaa
     3961 gtagagctgg aagtgcgtga cctccctgaa gagttgtctc tatccttcaa tgccacctgc
     4021 ctcaacaatg aggtcatccc tggcctcaag tcttgtatgg gactcaagat tggagacacg
     4081 gtgagcttca gcattgaggc caaggtgcga ggctgtcccc aggagaagga gaagtccttt
     4141 accataaagc ccgtgggctt caaggacagc ctgatcgtcc aggtcacctt tgattgtgac
     4201 tgtgcctgcc aggcccaagc tgaacctaat agccatcgct gcaacaatgg caatgggacc
     4261 tttgagtgtg gggtatgccg ttgtgggcct ggctggctgg gatcccagtg tgagtgctca
     4321 gaggaggact atcgcccttc ccagcaggac gaatgcagcc cccgggaggg tcagcccgtc
     4381 tgcagccagc ggggcgagtg cctctgtggt caatgtgtct gccacagcag tgactttggc
     4441 aagatcacgg gcaagtactg cgagtgtgac gacttctcct gtgtccgcta caagggggag
     4501 atgtgctcag gccatggcca gtgcagctgt ggggactgcc tgtgtgactc cgactggacc
     4561 ggctactact gcaactgtac cacgcgtact gacacctgca tgtccagcaa tgggctgctg
     4621 tgcagcggcc gcggcaagtg tgaatgtggc agctgtgtct gtatccagcc gggctcctat
     4681 ggggacacct gtgagaagtg ccccacctgc ccagatgcct gcacctttaa gaaagaatgt
     4741 gtggagtgta agaagtttga ccggggagcc ctacatgacg aaaatacctg caaccgttac
     4801 tgccgtgacg agattgagtc agtgaaagag cttaaggaca ctggcaagga tgcagtgaat
     4861 tgtacctata agaatgagga tgactgtgtc gtcagattcc agtactatga agattctagt
     4921 ggaaagtcca tcctgtatgt ggtagaagag ccagagtgtc ccaagggccc tgacatcctg
     4981 gtggtcctgc tctcagtgat gggggccatt ctgctcattg gccttgccgc cctgctcatc
     5041 tggaaactcc tcatcaccat ccacgaccga aaagaattcg ctaaatttga ggaagaacgc
     5101 gccagagcaa aatgggacac agccaacaac ccactgtata aagaggccac gtctaccttc
     5161 accaatatca cgtaccgggg cactgaattc gattacaagg atgacgacga taagtgagga
     5221 tcccgcgact ctagataatt ctaccgggta ggggaggcgc ttttcccaag gcagtctgga
     5281 gcatgcgctt tagcagcccc gctgggcact tggcgctaca caagtggcct ctggcctcgc
     5341 acacattcca catccaccgg taggcgccaa ccggctccgt tctttggtgg ccccttcgcg
     5401 ccaccttcta ctcctcccct agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc
     5461 aggacgtgac aaatggaagt agcacgtctc actagtctcg tgcagatgga cagcaccgct
     5521 gagcaatgga agcgggtagg cctttggggc agcggccaat agcagctttg ctccttcgct
     5581 ttctgggctc agaggctggg aaggggtggg tccgggggcg ggctcagggg cgggctcagg
     5641 ggcggggcgg gcgcccgaag gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc
     5701 acgtctgccg cgctgttctc ctcttcctca tctccgggcc tttcgacctg cagcccaagc
     5761 ttaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     5821 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     5881 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     5941 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     6001 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     6061 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     6121 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     6181 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     6241 tcctggagac ctccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     6301 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     6361 cctgaccgcg tctggaacaa tcaacctctg gattacaaaa tttgtgaaag attgactggt
     6421 attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat gcctttgtat
     6481 catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc ctggttgctg
     6541 tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg cactgtgttt
     6601 gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct ttccgggact
     6661 ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct tgcccgctgc
     6721 tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg gaagctgacg
     6781 tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac gtccttctgc
     6841 tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct gccggctctg
     6901 cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct ttgggccgcc
     6961 tccccgcctg gaattaattc tgcagtcgag acctagaaaa acatggagca atcacaagta
     7021 gcaatacagc agctaccaat gctgattgtg cctggctaga agcacaagag gaggaggagg
     7081 tgggttttcc agtcacacct caggtacctt taagaccaat gacttacaag gcagctgtag
     7141 atcttagcca ctttttaaaa gaaaagaggg gactggaagg gctaattcac tcccaacgaa
     7201 gacaagatat ccttgatctg tggatctacc acacacaagg ctacttccct gattagcaga
     7261 actacacacc agggccaggg gtcagatatc cactgacctt tggatggtgc tacaagctag
     7321 taccagttga gccagataag gtagaagagg ccaataaagg agagaacacc agcttgttac
     7381 accctgtgag cctgcatggg atggatgacc cggagagaga agtgttagag tggaggtttg
     7441 acagccgcct agcatttcat cacgtggccc gagagctgca tccggagtac ttcaagaact
     7501 gctgatatcg agcttgctac aagggacttt ccgctgggga ctttccaggg aggcgtggcc
     7561 tgggcgggac tggggagtgg cgagccctca gatcctgcat ataagcagct gctttttgcc
     7621 tgtactgggt ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg
     7681 aacccactgc ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt
     7741 ctgttgtgtg actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc
     7801 tctagcagta gtagttcatg tcatcttatt attcagtatt tataacttgc aaagaaatga
     7861 atatcagaga gtgagaggcc ttgacattgc tagcgtttta ccgtcgacct ctagctagag
     7921 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     7981 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     8041 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     8101 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     8161 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     8221 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     8281 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     8341 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     8401 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     8461 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     8521 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     8581 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     8641 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     8701 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     8761 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     8821 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     8881 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     8941 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     9001 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     9061 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     9121 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     9181 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     9241 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     9301 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     9361 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     9421 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     9481 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     9541 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     9601 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     9661 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     9721 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     9781 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     9841 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     9901 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     9961 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
    10021 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
    10081 gccacctgac gtcgacggat cgggagatca acttgtttat tgcagcttat aatggttaca
    10141 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
    10201 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gatcaactgg ataactcaag
    10261 ctaaccaaaa tcatcccaaa cttcccaccc cataccctat taccactgcc aattacctgt
    10321 ggtttcattt actctaaacc tgtgattcct ctgaattatt ttcattttaa agaaattgta
    10381 tttgttaaat atgtactaca aacttagtag tttttaaaga aattgtattt gttaaatatg
    10441 tactacaaac ttagtagt
//
