LOCUS       sequence-160465-        7592 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Can be used to produce inducible SIV-derived vector. Cells tranduced
            by lentiviral particles will overexpress GFP upon activation of the 
            tetO promoter. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7592)
  AUTHORS   Mangeot PE, Dollet S, Girard M, Ciancia C, Joly S, Peschanski M, 
            Lotteau V
  TITLE     Protein transfer into human cells by VSV-G-induced nanovesicles.
  JOURNAL   Mol Ther. 2011 Sep;19(9):1656-66. doi: 10.1038/mt.2011.138. Epub 
            2011 Jul 12.
  PUBMED    21750535
REFERENCE   2  (bases 1 to 7592)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/mt.2011.138"; journalName: "Mol Ther"; date: "2011-09"; 
            volume: "19"; issue: "9"; pages: "1656-66"
COMMENT     Sequence Label: GAE teo-GFP
FEATURES             Location/Qualifiers
     source          1..7592
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     588..610
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     602..619
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     603..619
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        764..1143
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1144..1347
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    join(2901..2919,2920..2942,2943..2961,2962..2984,2985..3003,
                     3004..3026,3027..3045,3046..3068,3069..3087,3088..3110,
                     3111..3129,3130..3152,3153..3171)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        3204..3242
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             join(3361..3363,3364..3366,3367..4080)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3361..3363,3364..3366,3367..4077)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3406..3427)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3667..3686)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4014..4035
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4087..4675
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4140..4160)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4558..4569)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     promoter        complement(5401..5419)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(5440..5456)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5440..5456)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5453..5475)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5464..5480
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5488..5494,5495..5512,5513..5518))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5533..5554
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5671..5688)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5842..6430)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5922..5941)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6601..7392,7393..7461))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7224..7243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7462..7566)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggaaattgta aacgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcg cgccattcgc cattcaggct
      481 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
      541 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
      601 ttgtaaaacg acggccagtg aattgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaattaattc ccattgcata cgttgtatcc atatcataat
      721 atgtacattt atattggctc atgtccaaca ttaccgccat gttgacattg attattgact
      781 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc
      841 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg
      901 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa
      961 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca
     1021 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac
     1081 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc
     1141 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga
     1201 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg
     1261 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta
     1321 cggtgggagg tctatataag cagagctcgt ttagtgaacc gcagtcgctc tgcggagagg
     1381 ctggcagatt gagccctggg aggttctctc cagcactagc aggtagagcc tgggtgttcc
     1441 ctgctagact ctcaccagca cttggccagt gctgggcaga gtggctccac gcttgcttgc
     1501 ttaaagacct cttcaataaa gctgccattt tagaagtaag ccagtgtgtg ttcccatctc
     1561 tcctagtcgc cgcctggtca actcggtact cggtaataag aagaccctgg tctgttagga
     1621 ccctttctgc tttgagaaac cgaagcagga aaatccctag cagattggcg cccgaacagg
     1681 acttgaagga gagtgagaga ctcctgagta cggctgagtg aaggcagtaa gggcggcagg
     1741 aaccaaccac gacggagtgc tcctataaag gcgcgggtcg gtaccagacg gcgtgaggag
     1801 cgggagagga ggaggcctcc ggttgcaggt aagtgcaaca caaaaaagaa atagctgtct
     1861 tgttatccag gaagggataa taagatagag tgggagatgg gcgcgagaaa ctccgtcttg
     1921 tcagggaaga aagcagatga attgaaaaaa ttattaatcg catgaatttt aaaagaaggg
     1981 gaggaatagg ggatatgact ccagcagaaa gattaattaa catgatcact acagaacaag
     2041 aaatacaatt tcaacaatca aaaaactcaa aatttaaaaa ttttcgggtc tattacagag
     2101 ctcacgcgtg attggagttg ggagattata aattagtaga gatcactccg attggcttgg
     2161 cccccacaga tgtgaagagg tacactactg gtggcacctc aagaaataaa agaggggtct
     2221 ttgtgctagg gttcttgggt tttctcgcaa cggcaggttc tgcaatgggc gcggcgtcgt
     2281 tgacgctgac cgctcagtcc cggactttat tggctgggat agtgcagcaa cagcaacagc
     2341 tgttggacgt ggtcaagaga caacaagaat tgttgcgact gaccgtctgg ggaacaaaga
     2401 acctccagac tagggtcact gccatcgaga agtacttaaa ggaccaggcg cagctaaatg
     2461 cttggggatg tgcgtttaga caagtctgcc acactactgt accatggcca aatgcaagtc
     2521 taacaccaga ctggaacaat gatacttggc aagagtggga gcgaaaggtt gacttcttgg
     2581 aggaaaatat aacagccctc ctagaagagg cacaaattca acaagagaag aacatgtatg
     2641 aattacaaaa gttgaatagc tgggatgtgt ttggcaattg gtttgacctt gcttcttgga
     2701 taaagtatat acaatatgga atttatgtag ttgtaggagt aatactgtta agaatagtga
     2761 tctatataga caaatgctag ctaagttaag gcaggggtat aggccagtgt tctcttcccc
     2821 caccctctta tttccatgat acgcagaccc aacagtaccc ttagcctatt atcgatccct
     2881 ttcgtccacg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
     2941 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
     3001 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
     3061 tttaccactc cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag
     3121 tgatagagaa aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa
     3181 gtcgagctcg gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag
     3241 ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt gacctccata
     3301 gaagacaccg ggaccgatcc agcctccgcg gccccggcta agctaccggt cgccaccagt
     3361 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3421 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3481 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3541 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3601 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3661 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3721 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3781 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3841 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3901 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3961 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     4021 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtga
     4081 ggatccaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4141 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca tgctattgct
     4201 tcccgtatgg ctttcatttt ctcctccttg tataaatcct ggttgctgtc tctttatgag
     4261 gagttgtggc ccgttgtcag gcaacgtggc gtggtgtgca ctgtgtttgc tgacgcaacc
     4321 cccactggtt ggggcattgc caccacctgt cagctccttt ccgggacttt cgctttcccc
     4381 ctccctattg ccacggcgga actcatcgcc gcctgccttg cccgctgctg gacaggggct
     4441 cggctgttgg gcactgacaa ttccgtggtg ttgtcgggga agctgacgtc ctttccatgg
     4501 ctgctcgcct gtgttgccac ctggattctg cgcgggacgt ccttctgcta cgtcccttcg
     4561 gccctcaatc cagcggacct tccttcccgc ggcctgctgc cggctctgcg gcctcttccg
     4621 cgtcttcgcc ttcgccctca gacgagtcgg atctcccttt gggccgcctc cccgcctgtt
     4681 tcgcctcgcg ataccgtcga gttttataaa agaaaagggg ggactggaag ggatttatta
     4741 cagtgcaaga agacatagaa tcttagacat gtacttagaa aaggaagaag gcatcatacc
     4801 agattggcag gattacacct caggaccagg aattagatac ccaaagacat ttggctggct
     4861 atggaaatta gtccctgtaa atgtatcaga tgaggcacag gaggatgaga ggcattattt
     4921 aatgcagcca gctcaaactt ccaagtggga tgacccttgg ggagaggttc tagcgtggaa
     4981 gtttgatcca actctagcct acacttatga ggcatatgct agatacccag aagagttgga
     5041 agcaagtcag gcctgtcaga actgcatttc gctctgtatt cagtcgctct gcggagaggc
     5101 tggcagattg agccctggga ggttctctcc agcactagca ggtagagcct gggtgttccc
     5161 tgctagactc tcaccagcac ttggccagtg ctgggcagag tggctccacg cttgcttgct
     5221 taaagacctc ttcaataaag ctgccatttt agaagtaagc cagtgtgtgt tcccatctct
     5281 cctagtcgcc gcctggtcaa ctcggtactc ggtaataaga agaccctggt ctgttaggac
     5341 cctttctgct ttgagaaacc gaagcaggaa aatccctagc atggtaccca gcttttgttc
     5401 cctttagtga gggttaattc cgagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5461 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5521 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5581 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5641 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     5701 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     5761 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     5821 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5881 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5941 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6001 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6061 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6121 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6181 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6241 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     6301 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6361 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6421 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6481 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6541 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6601 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6661 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6721 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6781 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6841 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6901 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6961 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7021 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7081 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7141 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7201 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7261 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7321 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7381 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7441 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7501 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7561 tccgcgcaca tttccccgaa aagtgccacc tg
//
