LOCUS       Exported               10021 bp DNA     circular SYN 28-JUL-2025
DEFINITION  Exported.
ACCESSION   V014892
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10021)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 10021)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10021
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             1..634
                     /label=3' LTR
                     /note="3' long terminal repeat (LTR) from HIV-1"
     misc_feature    681..806
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     misc_feature    1303..1536
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             1721..1765
                     /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             1914..1955
                     /label=Protein Tat
                     /note="Protein Tat from Human immunodeficiency virus type 1
                     group M subtype B (isolate WMJ22). Accession#: P12509"
     misc_feature    2027..2144
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     promoter        complement(2528..2895)
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     promoter        2912..3422
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3441..4184
                     /label=Tet-On(R) 3G
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of 
                     doxycycline"
     promoter        4198..4527
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4578..5243
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
     CDS             5313..5366
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             5376..5972
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     misc_feature    5989..6577
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     LTR             6784..7417
                     /label=5' LTR
                     /note="5' long terminal repeat (LTR) from HIV-1"
     promoter        complement(7594..7624)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7639..7660)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7948..8536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8710..9567)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9568..9672)
                     /label=AmpR promoter
     polyA_signal    9720..9854
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation 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 actacaaaaa caaattacaa aaattcaaaa ttttcgggtt tattacaggg
     2161 acagcagaga tccagtttat cgacttaact tgtttattgc agcttataat ggttacaaat
     2221 aaggcaatag catcacaaat ttcacaaata aggcattttt ttcactgcat tctagttttg
     2281 gtttgtccaa actcatcaat gtatcttatc atgtctggat ctcaaatccc tcggaagctg
     2341 cgcctgtctt aggttggagt gatacatttt tatcactttt acccgtcttt ggattaggca
     2401 gtagctctga cggccctcct gtcttaggtt agtgaaaaat gtcactctct tacccgtcat
     2461 tggctgtcca gcttagctcg caggggaggt ggtctggatc cgccggcacc ggtgtatacg
     2521 ggaattcttt acgagggtag gaagtggtac ggaaagttgg tataagacaa aagtgttgtg
     2581 gaattgaagt ttactcaaaa aatcagcact cttttatagg cgccctggtt tacataagca
     2641 aagcttatac gttctctatc actgataggg agtaaactgg atatacgttc tctatcactg
     2701 atagggagta aactgtagat acgttctcta tcactgatag ggagtaaact ggtcatacgt
     2761 tctctatcac tgatagggag taaactcctt atacgttctc tatcactgat agggagtaaa
     2821 gtctgcatac gttctctatc actgataggg agtaaactct tcatacgttc tctatcactg
     2881 atagggagta aactcgaggt gataattcca cggggttggg gttgcgcctt ttccaaggca
     2941 gccctgggtt tgcgcaggga cgcggctgct ctgggcgtgg ttccgggaaa cgcagcggcg
     3001 ccgaccctgg gtctcgcaca ttcttcacgt ccgttcgcag cgtcacccgg atcttcgccg
     3061 ctacccttgt gggccccccg gcgacgcttc ctgctccgcc cctaagtcgg gaaggttcct
     3121 tgcggttcgc ggcgtgccgg acgtgacaaa cggaagccgc acgtctcact agtaccctcg
     3181 cagacggaca gcgccaggga gcaatggcag cgcgccgacc gcgatgggct gtggccaata
     3241 gcggctgctc agcagggcgc gccgagagca gcggccggga aggggcggtg cgggaggcgg
     3301 ggtgtggggc ggtagtgtgg gccctgttcc tgcccgcgcg gtgttccgca ttctgcaagc
     3361 ctccggagcg cacgtcggca gtcggctccc tcgttgaccg aatcaccgac ctctctcccc
     3421 agggggatca tcgaattacc atgtctagac tggacaagag caaagtcata aactctgctc
     3481 tggaattact caatggagtc ggtatcgaag gcctgacgac aaggaaactc gctcaaaagc
     3541 tgggagttga gcagcctacc ctgtactggc acgtgaagaa caagcgggcc ctgctcgatg
     3601 ccctgccaat cgagatgctg gacaggcatc atacccactc ctgccccctg gaaggcgagt
     3661 catggcaaga ctttctgcgg aacaacgcca agtcataccg ctgtgctctc ctctcacatc
     3721 gcgacggggc taaagtgcat ctcggcaccc gcccaacaga gaaacagtac gaaaccctgg
     3781 aaaatcagct cgcgttcctg tgtcagcaag gcttctccct ggagaacgca ctgtacgctc
     3841 tgtccgccgt gggccacttt acactgggct gcgtattgga ggaacaggag catcaagtag
     3901 caaaagagga aagagagaca cctaccaccg attctatgcc cccacttctg aaacaagcaa
     3961 ttgagctgtt cgaccggcag ggagccgaac ctgccttcct tttcggcctg gaactaatca
     4021 tatgtggcct ggagaaacag ctaaagtgcg aaagcggcgg gccgaccgac gcccttgacg
     4081 attttgactt agacatgctc ccagccgatg cccttgacga ctttgacctt gatatgctgc
     4141 ctgctgacgc tcttgacgat tttgaccttg acatgctccc cgggtaaacg cgcgaatgtg
     4201 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4261 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     4321 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     4381 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     4441 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     4501 ttttttggag gcctaggctt ttgcaaatgt gaccggcgcc tacgctagac gccaccatgg
     4561 agagcgacga gagcggcctg cccgccatgg agatcgagtg ccgcatcacc ggcaccctga
     4621 acggcgtgga gttcgagctg gtgggcggcg gagagggcac ccccaagcag ggccgcatga
     4681 ccaacaagat gaagagcacc aaaggcgccc tgaccttcag cccctacctg ctgagccacg
     4741 tgatgggcta cggcttctac cacttcggca cctaccccag cggctacgag aaccccttcc
     4801 tgcacgccat caacaacggc ggctacacca acacccgcat cgagaagtac gaggacggcg
     4861 gcgtgctgca cgtgagcttc agctaccgct acgaggccgg ccgcgtgatc ggcgacttca
     4921 aggtggtggg caccggcttc cccgaggaca gcgtgatctt caccgacaag atcatccgca
     4981 gcaacgccac cgtggagcac ctgcacccca tgggcgataa cgtgctggtg ggcagcttcg
     5041 cccgcacctt cagcctgcgc gacggcggct actacagctt cgtggtggac agccacatgc
     5101 acttcaagag cgccatccac cccagcatcc tgcagaacgg gggccccatg ttcgccttcc
     5161 gccgcgtgga ggagctgcac agcaacaccg agctgggcat cgtggagtac cagcacgcct
     5221 tcaagacccc catcgccttc gccagatccc gcgctcagtc gtccaattct gccgtggacg
     5281 gcaccgccgg acccggctcc accggatctc gcgagggcag aggaagtctt ctaacatgcg
     5341 gtgacgtgga ggagaatccc ggccctgacg cgaccatgac cgagtacaag cccacggtgc
     5401 gcctcgccac ccgcgacgac gtcccccggg ccgtacgcac cctcgccgcc gcgttcgccg
     5461 actaccccgc cacgcgccac accgtcgacc cggaccgcca catcgagcgg gtcaccgagc
     5521 tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg
     5581 acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg
     5641 ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga
     5701 tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg
     5761 gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg
     5821 aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc
     5881 ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc
     5941 gcacctggtg catgacccgc aagcccggtg cctgaacgcg tctggaacaa tcaacctctg
     6001 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     6061 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     6121 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     6181 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     6241 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     6301 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     6361 aattccgtgg tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc
     6421 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     6481 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     6541 cagacgagtc ggatctccct ttgggccgcc tccccgcctg gaattaattc tgcagtcgag
     6601 acctagaaaa acatggagca atcacaagta gcaatacagc agctaccaat gctgattgtg
     6661 cctggctaga agcacaagag gaggaggagg tgggttttcc agtcacacct caggtacctt
     6721 taagaccaat gacttacaag gcagctgtag atcttagcca ctttttaaaa gaaaagaggg
     6781 gactggaagg gctaattcac tcccaacgaa gacaagatat ccttgatctg tggatctacc
     6841 acacacaagg ctacttccct gattagcaga actacacacc agggccaggg gtcagatatc
     6901 cactgacctt tggatggtgc tacaagctag taccagttga gccagataag gtagaagagg
     6961 ccaataaagg agagaacacc agcttgttac accctgtgag cctgcatggg atggatgacc
     7021 cggagagaga agtgttagag tggaggtttg acagccgcct agcatttcat cacgtggccc
     7081 gagagctgca tccggagtac ttcaagaact gctgatatcg agcttgctac aagggacttt
     7141 ccgctgggga ctttccaggg aggcgtggcc tgggcgggac tggggagtgg cgagccctca
     7201 gatcctgcat ataagcagct gctttttgcc tgtactgggt ctctctggtt agaccagatc
     7261 tgagcctggg agctctctgg ctaactaggg aacccactgc ttaagcctca ataaagcttg
     7321 ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg actctggtaa ctagagatcc
     7381 ctcagaccct tttagtcagt gtggaaaatc tctagcagta gtagttcatg tcatcttatt
     7441 attcagtatt tataacttgc aaagaaatga atatcagaga gtgagaggcc ttgacattgc
     7501 tagcgtttta ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     7561 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     7621 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     7681 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     7741 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     7801 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     7861 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     7921 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     7981 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     8041 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     8101 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     8161 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     8221 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     8281 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     8341 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg
     8401 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     8461 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     8521 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     8581 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     8641 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     8701 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8761 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8821 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     8881 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8941 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     9001 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     9061 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     9121 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     9181 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     9241 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     9301 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     9361 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     9421 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     9481 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     9541 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     9601 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     9661 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat cgggagatca
     9721 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     9781 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     9841 atcatgtctg gatcaactgg ataactcaag ctaaccaaaa tcatcccaaa cttcccaccc
     9901 cataccctat taccactgcc aattacctgt ggtttcattt actctaaacc tgtgattcct
     9961 ctgaattatt ttcattttaa agaaattgta tttgttaaat atgtactaca aacttagtag
    10021 t
//
