LOCUS       V019536                 8949 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V019536
VERSION     V019536
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             2830..2856
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2857..3705
                     /label="MTAP(NM_002451)"
                     /note="MTAP(NM_002451)"
                     /gene="MTAP"
     promoter        3737..4236
                     /note="mouse phosphoglycerate kinase 1 promoter"
                     /label="PGK promoter"
     CDS             4257..4856
                     /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    4870..5458
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     LTR             5665..6298
                     /note="3' long terminal repeat (LTR) from HIV-1"
                     /label="3' LTR"
     primer_bind     complement(6427..6443)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6451..6467
                     /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(6475..6505)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6520..6541
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6829..7417)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7588..8448)
                     /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(8449..8553)
                     /gene="bla"
                     /label="AmpR promoter"
     polyA_signal    8601..8735
                     /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 gccaccatgt acccatacga cgtaccagat tacgctatgg cctctggcac caccaccacc
     2881 gccgtgaaga ttggaataat tggtggaaca ggcctggatg atccagaaat tttagaagga
     2941 agaactgaaa aatatgtgga tactccattt ggcaagccat ctgatgcctt aattttgggg
     3001 aagataaaaa atgttgattg cgtcctcctt gcaaggcatg gaaggcagca caccatcatg
     3061 ccttcaaagg tcaactacca ggcgaacatc tgggctttga aggaagaggg ctgtacacat
     3121 gtcatagtga ccacagcttg tggctccttg agggaggaga ttcagcccgg cgatattgtc
     3181 attattgatc agttcattga caggaccact atgagacctc agtccttcta tgatggaagt
     3241 cattcttgtg ccagaggagt gtgccatatt ccaatggctg agccgttttg ccccaaaacg
     3301 agagaggttc ttatagagac tgctaagaag ctaggactcc ggtgccactc aaaggggaca
     3361 atggtcacaa tcgagggacc tcgttttagc tcccgggcag aaagcttcat gttccgcacc
     3421 tggggggcgg atgttatcaa catgaccaca gttccagagg tggttcttgc taaggaggct
     3481 ggaatttgtt acgcaagtat cgccatggcg acagattatg actgctggaa ggagcacgag
     3541 gaagcagttt cggtggaccg ggtcttaaag accctgaaag aaaacgctaa taaagccaaa
     3601 agcttactgc tcactaccat acctcagata gggtccacag aatggtcaga aaccctccat
     3661 aacctgaaga atatggccca gttttctgtt ttattaccaa gacattaagg atcccgcgac
     3721 tctagataat tctaccgggt aggggaggcg cttttcccaa ggcagtctgg agcatgcgct
     3781 ttagcagccc cgctgggcac ttggcgctac acaagtggcc tctggcctcg cacacattcc
     3841 acatccaccg gtaggcgcca accggctccg ttctttggtg gccccttcgc gccaccttct
     3901 actcctcccc tagtcaggaa gttccccccc gccccgcagc tcgcgtcgtg caggacgtga
     3961 caaatggaag tagcacgtct cactagtctc gtgcagatgg acagcaccgc tgagcaatgg
     4021 aagcgggtag gcctttgggg cagcggccaa tagcagcttt gctccttcgc tttctgggct
     4081 cagaggctgg gaaggggtgg gtccgggggc gggctcaggg gcgggctcag gggcggggcg
     4141 ggcgcccgaa ggtcctccgg aggcccggca ttctgcacgc ttcaaaagcg cacgtctgcc
     4201 gcgctgttct cctcttcctc atctccgggc ctttcgacct gcagcccaag cttaccatga
     4261 ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca
     4321 ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc
     4381 acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg
     4441 gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg
     4501 tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc
     4561 ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg
     4621 cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg
     4681 ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga
     4741 cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg
     4801 tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgaccgc
     4861 gtctggaaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     4921 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     4981 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     5041 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     5101 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     5161 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     5221 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     5281 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     5341 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     5401 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     5461 ggaattaatt ctgcagtcga gacctagaaa aacatggagc aatcacaagt agcaatacag
     5521 cagctaccaa tgctgattgt gcctggctag aagcacaaga ggaggaggag gtgggttttc
     5581 cagtcacacc tcaggtacct ttaagaccaa tgacttacaa ggcagctgta gatcttagcc
     5641 actttttaaa agaaaagagg ggactggaag ggctaattca ctcccaacga agacaagata
     5701 tccttgatct gtggatctac cacacacaag gctacttccc tgattagcag aactacacac
     5761 cagggccagg ggtcagatat ccactgacct ttggatggtg ctacaagcta gtaccagttg
     5821 agccagataa ggtagaagag gccaataaag gagagaacac cagcttgtta caccctgtga
     5881 gcctgcatgg gatggatgac ccggagagag aagtgttaga gtggaggttt gacagccgcc
     5941 tagcatttca tcacgtggcc cgagagctgc atccggagta cttcaagaac tgctgatatc
     6001 gagcttgcta caagggactt tccgctgggg actttccagg gaggcgtggc ctgggcggga
     6061 ctggggagtg gcgagccctc agatcctgca tataagcagc tgctttttgc ctgtactggg
     6121 tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg
     6181 cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt
     6241 gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt
     6301 agtagttcat gtcatcttat tattcagtat ttataacttg caaagaaatg aatatcagag
     6361 agtgagaggc cttgacattg ctagcgtttt accgtcgacc tctagctaga gcttggcgta
     6421 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     6481 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     6541 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6601 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     6661 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     6721 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     6781 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     6841 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     6901 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     6961 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     7021 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     7081 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     7141 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     7201 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     7261 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     7321 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     7381 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     7441 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     7501 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     7561 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     7621 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     7681 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     7741 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     7801 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     7861 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     7921 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     7981 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     8041 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     8101 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     8161 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     8221 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     8281 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     8341 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     8401 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     8461 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     8521 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     8581 cgtcgacgga tcgggagatc aacttgttta ttgcagctta taatggttac aaataaagca
     8641 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     8701 ccaaactcat caatgtatct tatcatgtct ggatcaactg gataactcaa gctaaccaaa
     8761 atcatcccaa acttcccacc ccatacccta ttaccactgc caattacctg tggtttcatt
     8821 tactctaaac ctgtgattcc tctgaattat tttcatttta aagaaattgt atttgttaaa
     8881 tatgtactac aaacttagta gtttttaaag aaattgtatt tgttaaatat gtactacaaa
     8941 cttagtagt
//
