LOCUS       V014337                 8967 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014337
VERSION     V014337
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 8967)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..131
                     /label="AmpR promoter"
     CDS             132..989
                     /label="AmpR"
                     /note="beta-lactamase"
     rep_origin      1163..1751
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     protein_bind    2039..2060
                     /label="CAP binding site"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2075..2105
                     /label="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    2113..2129
                     /label="lac operator"
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     2137..2153
                     /label="M13 rev"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     promoter        2174..2192
                     /label="T3 promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2220..2447
                     /label="RSV promoter"
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2448..2628
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2672..2797
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    3290..3523
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             3707..3751
                     /label="gp41 peptide"
                     /note="antigenic peptide corresponding to amino acids 655
                     to 669 of the HIV envelope protein gp41 (Lutje Hulsik et
                     al., 2013)"
     CDS             3900..3941
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    4012..4129
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     enhancer        4152..4455
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4456..4659
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             4832..5788
                     /label="TurboID"
                     /note="mutant of the E. coli biotin ligase BioID engineered
                     for improved catalytic efficiency (Branon et al., 2018)"
     CDS             5795..5815
                     /label="SV40 NLS"
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     CDS             5816..5839
                     /label="FLAG"
                     /note="FLAG(R) epitope tag, followed by an enterokinase
                     cleavage site"
     misc_feature    5849..6437
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     promoter        6451..6950
                     /label="PGK promoter"
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             6971..7567
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     LTR             7708..7941
                     /label="3' LTR (Delta-U3)"
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    8013..8147
                     /label="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      8153..8288
                     /label="SV40 ori"
                     /note="SV40 origin of replication"
     promoter        complement(8321..8339)
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(8349..8365)
                     /label="M13 fwd"
                     /note="common sequencing primer, one of multiple similar
                     variants"
     rep_origin      8510..8965
                     /direction=RIGHT
                     /label="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
       61 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      121 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      181 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      241 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      301 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
      361 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
      421 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
      481 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
      541 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
      601 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
      661 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
      721 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
      781 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
      841 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
      901 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
      961 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1021 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1081 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1141 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1201 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1261 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     1321 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     1381 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     1441 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     1501 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     1561 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     1621 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     1681 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     1741 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     1801 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     1861 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     1921 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     1981 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2041 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2101 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2161 acgccaagcg cgcaattaac cctcactaaa gggaacaaaa gctggagctg caagcttaat
     2221 gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt agcaacatgc
     2281 cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt ggtacgatcg
     2341 tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca ctgaattgcc
     2401 gcattgcaga gatattgtat ttaagtgcct agctcgatac aataaacggg tctctctggt
     2461 tagaccagat ctgagcctgg gagctctctg gctaactagg gaacccactg cttaagcctc
     2521 aataaagctt gccttgagtg cttcaagtag tgtgtgcccg tctgttgtgt gactctggta
     2581 actagagatc cctcagaccc ttttagtcag tgtggaaaat ctctagcagt ggcgcccgaa
     2641 cagggacctg aaagcgaaag ggaaaccaga gctctctcga cgcaggactc ggcttgctga
     2701 agcgcgcacg gcaagaggcg aggggcggcg actggtgagt acgccaaaaa ttttgactag
     2761 cggaggctag aaggagagag atgggtgcga gagcgtcagt attaagcggg ggagaattag
     2821 atcgcgatgg gaaaaaattc ggttaaggcc agggggaaag aaaaaatata aattaaaaca
     2881 tatagtatgg gcaagcaggg agctagaacg attcgcagtt aatcctggcc tgttagaaac
     2941 atcagaaggc tgtagacaaa tactgggaca gctacaacca tcccttcaga caggatcaga
     3001 agaacttaga tcattatata atacagtagc aaccctctat tgtgtgcatc aaaggataga
     3061 gataaaagac accaaggaag ctttagacaa gatagaggaa gagcaaaaca aaagtaagac
     3121 caccgcacag caagcggccg ctgatcttca gacctggagg aggagatatg agggacaatt
     3181 ggagaagtga attatataaa tataaagtag taaaaattga accattagga gtagcaccca
     3241 ccaaggcaaa gagaagagtg gtgcagagag aaaaaagagc agtgggaata ggagctttgt
     3301 tccttgggtt cttgggagca gcaggaagca ctatgggcgc agcctcaatg acgctgacgg
     3361 tacaggccag acaattattg tctggtatag tgcagcagca gaacaatttg ctgagggcta
     3421 ttgaggcgca acagcatctg ttgcaactca cagtctgggg catcaagcag ctccaggcaa
     3481 gaatcctggc tgtggaaaga tacctaaagg atcaacagct cctggggatt tggggttgct
     3541 ctggaaaact catttgcacc actgctgtgc cttggaatgc tagttggagt aataaatctc
     3601 tggaacagat tggaatcaca cgacctggat ggagtgggac agagaaatta acaattacac
     3661 aagcttaata cactccttaa ttgaagaatc gcaaaaccag caagaaaaga atgaacaaga
     3721 attattggaa ttagataaat gggcaagttt gtggaattgg tttaacataa caaattggct
     3781 gtggtatata aaattattca taatgatagt aggaggcttg gtaggtttaa gaatagtttt
     3841 tgctgtactt tctatagtga atagagttag gcagggatat tcaccattat cgtttcagac
     3901 ccacctccca accccgaggg gacccgacag gcccgaagga atagaagaag aaggtggaga
     3961 gagagacaga gacagatcca ttcgattagt gaacggatct cgacggttaa cttttaaaag
     4021 aaaagggggg attggggggt acagtgcagg ggaaagaata gtagacataa tagcaacaga
     4081 catacaaact aaagaattac aaaaacaaat tacaaaaatt caaaatttta tcgataagct
     4141 tgggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4201 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4261 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4321 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4381 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     4441 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     4501 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4561 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     4621 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagatc
     4681 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccgact ctagaggatc
     4741 cgtatacgta taccccgggg atatcacgcg tatgtgtaca ggcggcggcg gcagcggcgg
     4801 cggcggcagc ggcggcggcg gcagcgctag caaagacaat actgtgcctc tgaagctgat
     4861 cgctctcctg gctaatggcg agttccatag tggcgaacag ctgggagaaa ccctgggcat
     4921 gtccagggcc gctatcaaca agcacattca gactctgcgc gactggggcg tggacgtgtt
     4981 caccgtgccc ggaaagggct actctctgcc cgagcctatc ccgctgctga acgctaaaca
     5041 gattctggga cagctggacg gcgggagcgt ggcagtcctg cctgtggtcg actccaccaa
     5101 tcagtacctg ctggatcgaa tcggcgagct gaagagtggg gatgcttgca ttgcagaata
     5161 tcagcaggca gggagaggaa gcagagggag gaaatggttc tctccttttg gagctaacct
     5221 gtacctgagt atgttttggc gcctgaagcg gggaccagca gcaatcggcc tgggcccggt
     5281 catcggaatt gtcatggcag aagcgctgcg aaagctggga gcagacaagg tgcgagtcaa
     5341 atggcccaat gacctgtatc tgcaggatag aaagctggca ggcatcctgg tggagctggc
     5401 cggaataaca ggcgatgctg cacagatcgt cattggcgcc gggattaacg tggctatgag
     5461 gcgcgtggag gaaagcgtgg tcaatcaggg ctggatcaca ctgcaggaag cagggattaa
     5521 cctggacagg aatactctgg ccgctacgct gatccgagag ctgcgggcag ccctggaact
     5581 gttcgagcag gaaggcctgg ctccatatct gccacggtgg gagaagctgg ataacttcat
     5641 caatagaccc gtgaagctga tcattgggga caaagagatt ttcgggatta gccgggggat
     5701 tgataaacag ggagccctgc tgctggaaca ggacggagtt atcaaaccct ggatgggcgg
     5761 agaaatcagt ctgcggtctg ccgaaaagct gcagcccaag aagaagagga aggtggacta
     5821 caaggatgac gacgataagt aagtcgacaa tcaacctctg gattacaaaa tttgtgaaag
     5881 attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg ctgctttaat
     5941 gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct tgtataaatc
     6001 ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg gcgtggtgtg
     6061 cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct gtcagctcct
     6121 ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg ccgcctgcct
     6181 tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg tgttgtcggg
     6241 gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc tgcgcgggac
     6301 gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc gcggcctgct
     6361 gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc ggatctccct
     6421 ttgggccgcc tccccgcctg gaattctacc gggtagggga ggcgcttttc ccaaggcagt
     6481 ctggagcatg cgctttagca gccccgctgg gcacttggcg ctacacaagt ggcctctggc
     6541 ctcgcacaca ttccacatcc accggtaggc gccaaccggc tccgttcttt ggtggcccct
     6601 tcgcgccacc ttctactcct cccctagtca ggaagttccc ccccgccccg cagctcgcgt
     6661 cgtgcaggac gtgacaaatg gaagtagcac gtctcactag tctcgtgcag atggacagca
     6721 ccgctgagca atggaagcgg gtaggccttt ggggcagcgg ccaatagcag ctttgctcct
     6781 tcgctttctg ggctcagagg ctgggaaggg gtgggtccgg gggcgggctc aggggcgggc
     6841 tcaggggcgg ggcgggcgcc cgaaggtcct ccggaggccc ggcattctgc acgcttcaaa
     6901 agcgcacgtc tgccgcgctg ttctcctctt cctcatctcc gggcctttcg acctgcagcc
     6961 caagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     7021 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     7081 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     7141 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     7201 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     7261 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     7321 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     7381 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     7441 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     7501 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     7561 cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
     7621 tgcatctcga gggcccggta cctttaagac caatgactta caaggcagct gtagatctta
     7681 gccacttttt aaaagaaaag gggggactgg aagggctagc tcactcccaa cgaagacaag
     7741 atctgctttt tgcttgtact gggtctctct ggttagacca gatctgagcc tgggagctct
     7801 ctggctgcct agggaaccca ctgcttaagc ctcaataaag cttgccttga gtgcttcaag
     7861 tagtgtgtgc ccgtctgttg tgtgactctg gtaactagag atccctcaga cccttttagt
     7921 cagtgtggaa aatctctagc agtagtagtt catgtcatct tattattcag tatttataac
     7981 ttgcaaagaa atgaatatca gagagtgaga ggaacttgtt tattgcagct tataatggtt
     8041 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     8101 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggctctag ctatcccgcc
     8161 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     8221 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     8281 tggaggccta ggcttttgcg tcgagacgta cccaattcgc cctatagtga gtcgtattac
     8341 gcgcgctcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     8401 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     8461 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcga cgcgccctgt
     8521 agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc
     8581 agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac gttcgccggc
     8641 tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag tgctttacgg
     8701 cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc atcgccctga
     8761 tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg actcttgttc
     8821 caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata agggattttg
     8881 ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt
     8941 aacaaaatat taacgtttac aatttcc
//
