LOCUS       V014475                10709 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  Exported.
ACCESSION   V014475
VERSION     V014475
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
            .
REFERENCE   1  (bases 1 to 10709)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..10709
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        237..616
                     /label="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..819
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     misc_feature    1061..1186
                     /label="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1683..1916
                     /label="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for
                     Rev-dependent mRNA export from the nucleus to the
                     cytoplasm."
     CDS             2101..2145
                     /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             2294..2335
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
                     /label="Protein Tat"
     misc_feature    2443..2560
                     /label="cPPT/CTS"
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     protein_bind    2619..2639
                     /label="attB4"
                     /note="core recombination site for the Gateway(R) BP
                     reaction"
     promoter        2640..3818
                     /label="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     protein_bind    3818..3842
                     /label="attB1"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4390..4414)
                     /label="attB2"
                     /note="recombination site for the Gateway(R) BP reaction"
     promoter        4817..5016
                     /label="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        5045..5063
                     /label="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5084..6733
                     /label="luciferase"
                     /note="firefly luciferase"
     CDS             6803..6856
                     /label="T2A"
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             6857..7453
                     /label="PuroR"
                     /note="puromycin N-acetyltransferase"
     protein_bind    complement(7509..7542)
                     /label="loxP"
                     /note="Cre-mediated recombination occurs in the 8-bp core
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    7598..8186
                     /label="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             8715..8895
                     /label="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      complement(8957..9545)
                     /direction=LEFT
                     /label="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
     CDS             complement(9719..10576)
                     /label="AmpR"
                     /note="beta-lactamase"
     promoter        complement(10577..10681)
                     /label="AmpR promoter"
ORIGIN
        1 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
       61 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
      121 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      181 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgctgatct tcagacctgg aggaggagat
     1561 atgagggaca attggagaag tgaattatat aaatataaag tagtaaaaat tgaaccatta
     1621 ggagtagcac ccaccaaggc aaagagaaga gtggtgcaga gagaaaaaag agcagtggga
     1681 ataggagctt tgttccttgg gttcttggga gcagcaggaa gcactatggg cgcagcgtca
     1741 atgacgctga cggtacaggc cagacaatta ttgtctggta tagtgcagca gcagaacaat
     1801 ttgctgaggg ctattgaggc gcaacagcat ctgttgcaac tcacagtctg gggcatcaag
     1861 cagctccagg caagaatcct ggctgtggaa agatacctaa aggatcaaca gctcctgggg
     1921 atttggggtt gctctggaaa actcatttgc accactgctg tgccttggaa tgctagttgg
     1981 agtaataaat ctctggaaca gatttggaat cacacgacct ggatggagtg ggacagagaa
     2041 attaacaatt acacaagctt aatacactcc ttaattgaag aatcgcaaaa ccagcaagaa
     2101 aagaatgaac aagaattatt ggaattagat aaatgggcaa gtttgtggaa ttggtttaac
     2161 ataacaaatt ggctgtggta tataaaatta ttcataatga tagtaggagg cttggtaggt
     2221 ttaagaatag tttttgctgt actttctata gtgaatagag ttaggcaggg atattcacca
     2281 ttatcgtttc agacccacct cccaaccccg aggggacccg acaggcccga aggaatagaa
     2341 gaagaaggtg gagagagaga cagagacaga tccattcgat tagtgaacgg atcggcactg
     2401 cgtgcgccaa ttctgcagac aaatggcagt attcatccac aattttaaaa gaaaaggggg
     2461 gattgggggg tacagtgcag gggaaagaat agtagacata atagcaacag acatacaaac
     2521 taaagaatta caaaaacaaa ttacaaaaat tcaaaatttt cgggtttatt acagggacag
     2581 cagagatcca gtttggttag taccgggccc gctctagaca actttgtata gaaaagttgg
     2641 gctccggtgc ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttgtggg
     2701 gaggggtcgg caattgaacc ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg
     2761 atgtcgtgta ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag
     2821 tagtcgccgt gaacgttctt tttcgcaacg ggtttgccgc cagaacacag gtaagtgccg
     2881 tgtgtggttc ccgcgggcct ggcctcttta cgggttatgg cccttgcgtg ccttgaatta
     2941 cttccacctg gctgcagtac gtgattcttg atcccgagct tcgggttgga agtgggtggg
     3001 agagttcgag gccttgcgct taaggagccc cttcgcctcg tgcttgagtt gaggcctggc
     3061 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
     3121 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
     3181 aagatagtct tgtaaatgcg ggccaagatc tgcacactgg tatttcggtt tttggggccg
     3241 cgggcggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
     3301 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
     3361 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
     3421 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctgc agggagctca aaatggagga
     3481 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
     3541 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
     3601 agttctcgag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     3661 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     3721 tctccttgga atttgccctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     3781 tggttcaaag tttttttctt ccatttcagg tgtcgtgaca agtttgtaca aaaaagcagg
     3841 ctaatcttct ctcacctcct agacacacac acacagaggc acctccggga ggctctggcc
     3901 aggcctgggt gtggcaggat agaaatgaaa gttgtcctgg caacacacaa gatttcatca
     3961 acatttccat ccccagcagc tcctgggcca agcctcatct tgcccagtgt ggcctgtcaa
     4021 gatgacggcc tggaagagaa gccagactgg gcagccaggc ctctgcccct ccccttgcct
     4081 ggtggcacag ccagcactgg ctcactggtg agttgactaa tttcctccgc caccattagt
     4141 tgtccatgtt cattgctgcc tgacacccaa caccagtctg caagggcacc aggaggggat
     4201 ggaagagaca gggtgtctgg ggaagggtgg gtctgagtgc gagggggtgg gtgagcttgt
     4261 ctcagagcct gacgctggct ggtggtatct gcgttaccca ggctgaccca gctctctctg
     4321 atgcagacag gcgagggcct cacacacttc agagaattca cacccacatg atgaggctga
     4381 gaggagcaga cccagctttc ttgtacaaag tggtgataat cgaattggat ccgttgacat
     4441 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
     4501 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     4561 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     4621 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     4681 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     4741 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
     4801 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
     4861 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     4921 caaaatcaac gggactttcc aaaatgtcgt aataaccccg ccccgttgac gcaaatgggc
     4981 ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa ccgtcagaat
     5041 tttgtaatac gactcactat agggcggccg ggaattcgcc accatggaag acgccaaaaa
     5101 cataaagaaa ggcccggcgc cattctatcc gctggaagat ggaaccgctg gagagcaact
     5161 gcataaggct atgaagagat acgccctggt tcctggaaca attgctttta cagatgcaca
     5221 tatcgaggtg gacatcactt acgctgagta cttcgaaatg tccgttcggt tggcagaagc
     5281 tatgaaacga tatgggctga atacaaatca cagaatcgtc gtatgcagtg aaaactctct
     5341 tcaattcttt atgccggtgt tgggcgcgtt atttatcgga gttgcagttg cgcccgcgaa
     5401 cgacatttat aatgaacgtg aattgctcaa cagtatgggc atttcgcagc ctaccgtggt
     5461 gttcgtttcc aaaaaggggt tgcaaaaaat tttgaacgtg caaaaaaagc tcccaatcat
     5521 ccaaaaaatt attatcatgg attctaaaac ggattaccag ggatttcagt cgatgtacac
     5581 gttcgtcaca tctcatctac ctcccggttt taatgaatac gattttgtgc cagagtcctt
     5641 cgatagggac aagacaattg cactgatcat gaactcctct ggatctactg gtctgcctaa
     5701 aggtgtcgct ctgcctcata gaactgcctg cgtgagattc tcgcatgcca gagatcctat
     5761 ttttggcaat caaatcattc cggatactgc gattttaagt gttgttccat tccatcacgg
     5821 ttttggaatg tttactacac tcggatattt gatatgtgga tttcgagtcg tcttaatgta
     5881 tagatttgaa gaagagctgt ttctgaggag ccttcaggat tacaagattc aaagtgcgct
     5941 gctggtgcca accctattct ccttcttcgc caaaagcact ctgattgaca aatacgattt
     6001 atctaattta cacgaaattg cttctggtgg cgctcccctc tctaaggaag tcggggaagc
     6061 ggttgccaag aggttccatc tgccaggtat caggcaagga tatgggctca ctgagactac
     6121 atcagctatt ctgattacac ccgaggggga tgataaaccg ggcgcggtcg gtaaagttgt
     6181 tccatttttt gaagcgaagg ttgtggatct ggataccggg aaaacgctgg gcgttaatca
     6241 aagaggcgaa ctgtgtgtga gaggtcctat gattatgtcc ggttatgtaa acaatccgga
     6301 agcgaccaac gccttgattg acaaggatgg atggctacat tctggagaca tagcttactg
     6361 ggacgaagac gaacacttct tcatcgttga ccgcctgaag tctctgatta agtacaaagg
     6421 ctatcaggtg gctcccgctg aattggaatc catcttgctc caacacccca acatcttcga
     6481 cgcaggtgtc gcaggtcttc ccgacgatga cgccggtgaa cttcccgccg ccgttgttgt
     6541 tttggagcac ggaaagacga tgacggaaaa agagatcgtg gattacgtcg ccagtcaagt
     6601 aacaaccgcg aaaaagttgc gcggaggagt tgtgtttgtg gacgaagtac cgaaaggtct
     6661 taccggaaaa ctcgacgcaa gaaaaatcag agagatcctc ataaaggcca agaagggcgg
     6721 aaagatcgcc gtgagatccc gcgctcagtc gtccaattct gccgtggacg gcaccgccgg
     6781 acccggctcc accggatctc gcgagggcag aggaagtctt ctaacatgcg gtgacgtgga
     6841 ggagaatccc ggccctatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga
     6901 cgtcccccgg gcagtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca
     6961 caccgtcgat ccagaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac
     7021 gcgcgtcggg ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt
     7081 ctggaccacg ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat
     7141 ggccgagttg agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc
     7201 gcaccggccc aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca
     7261 gggcaagggt ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg
     7321 ggtgcccgcc ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg
     7381 cttcaccgtc accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg
     7441 caagcccggt gcctgagttt aaacgggccg gccgcggtct gtacaagtag gattcgtcga
     7501 gggacctaat aacttcgtat agcatacatt atacgaagtt atacatgttt aagggttccg
     7561 gttccactag gtacaattcg atatcaagct tatcgataat caacctctgg attacaaaat
     7621 ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat gtggatacgc
     7681 tgctttaatg cctttgtatc atgctattgc ttcccgtatg gctttcattt tctcctcctt
     7741 gtataaatcc tggttgctgt ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg
     7801 cgtggtgtgc actgtgtttg ctgacgcaac ccccactggt tggggcattg ccaccacctg
     7861 tcagctcctt tccgggactt tcgctttccc cctccctatt gccacggcgg aactcatcgc
     7921 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     7981 gttgtcgggg aaatcatcgt cctttccttg gctgctcgcc tgtgttgcca cctggattct
     8041 gcgcgggacg tccttctgct acgtcccttc ggccctcaat ccagcggacc ttccttcccg
     8101 cggcctgctg ccggctctgc ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg
     8161 gatctccctt tgggccgcct ccccgcatcg ataccgtcga cctcgatcga gacctagaaa
     8221 aacatggagc aatcacaagt agcaatacag cagctaccaa tgctgattgt gcctggctag
     8281 aagcacaaga ggaggaggag gtgggttttc cagtcacacc tcaggtacct ttaagaccaa
     8341 tgacttacaa ggcagctgta gatcttagcc actttttaaa agaaaagggg ggactggaag
     8401 ggctaattca ctcccaacga agacaagata tccttgatct gtggatctac cacacacaag
     8461 gctacttccc tgattggcag aactacacac cagggccagg gatcagatat ccactgacct
     8521 ttggatggtg ctacaagcta gtaccagttg agcaagagaa ggtagaagaa gccaatgaag
     8581 gagagaacac ccgcttgtta caccctgtga gcctgcatgg gatggatgac ccggagagag
     8641 aagtattaga gtggaggttt gacagccgcc tagcatttca tcacatggcc cgagagctgc
     8701 atccggactg tactgggtct ctctggttag accagatctg agcctgggag ctctctggct
     8761 aactagggaa cccactgctt aagcctcaat aaagcttgcc ttgagtgctt caagtagtgt
     8821 gtgcccgtct gttgtgtgac tctggtaact agagatccct cagacccttt tagtcagtgt
     8881 ggaaaatctc tagcagcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     8941 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     9001 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     9061 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     9121 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     9181 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     9241 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     9301 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     9361 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     9421 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     9481 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     9541 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     9601 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     9661 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     9721 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     9781 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     9841 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     9901 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     9961 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
    10021 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
    10081 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
    10141 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
    10201 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
    10261 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
    10321 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
    10381 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
    10441 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
    10501 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
    10561 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
    10621 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggtcccgc
    10681 gcacatttcc ccgaaaagtg ccacctgac
//
