LOCUS       sequence-101864-        7882 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Self-inactivating retrovirus vector containing the GFP gene.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7882)
  AUTHORS   Fujita T, Fujii H
  TITLE     Identification of Proteins Associated with an IFNgamma-Responsive 
            Promoter by a Retroviral Expression System for enChIP Using CRISPR.
  JOURNAL   PLoS One. 2014 Jul 22;9(7):e103084. doi: 
            10.1371/journal.pone.0103084. eCollection 2014.
  PUBMED    25051498
REFERENCE   2  (bases 1 to 7882)
  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"; journalName: "PLoS One.";
            date: "2014-07-22"; volume: "9(7):e103084. doi"; pages: " 
            10.1371/journal.pone.0103084. eCollection 2014"
COMMENT     Sequence Label: pSIR-GFP
FEATURES             Location/Qualifiers
     source          1..7882
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    580..937
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             996..1412
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1342..1364
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1380..1396
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      2814..2823
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2820..2822,2823..2825,2826..3539)
                     /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(2820..2822,2823..2825,2826..3536)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2865..2886)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3126..3145)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3473..3494
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4866..4882)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4866..4882)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4879..4901)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4890..4906
                     /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(4914..4920,4921..4938,4939..4944))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4959..4980
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5097..5114)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5268..5856)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5348..5367)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6027..6818,6819..6887))
                     /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     6650..6669
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6888..6992)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7060..7078
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7116..7138)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7238..7257
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     7451..7473
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7465..7482
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7466..7482
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
       61 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
      121 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
      181 gaacagatgg tccccagatg cggtccagcc ctcagcagtt tctagagaac catcagatgt
      241 ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt atttgaacta accaatcagt
      301 tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc tcaataaaag agcccacaac
      361 ccctcactcg gggcgccagt cctccgattg actgagtcgc ccgggtaccc gtgtatccaa
      421 taaaccctct tgcagttgca tccgacttgt ggtctcgctg ttccttggga gggtctcctc
      481 tgagtgattg actacccgtc agcgggggtc tttcatttgg gggctcgtcc gggatcggga
      541 gacccctgcc cagggaccac cgacccacca ccgggaggta agctggccag caacttatct
      601 gtgtctgtcc gattgtctag tgtctatgac tgattttatg cgcctgcgtc ggtactagtt
      661 agctaactag ctctgtatct ggcggacccg tggtggaact gacgagttcg gaacacccgg
      721 ccgcaaccct gggagacgtc ccagggactt cgggggccgt ttttgtggcc cgacctgagt
      781 ccaaaaatcc cgatcgtttt ggactctttg gtgcaccccc cttagaggag ggatatgtgg
      841 ttctggtagg agacgagaac ctaaaacagt tcccgcctcc gtctgaattt ttgctttcgg
      901 tttgggaccg aagccgcgcc gcgcgtcttg tctgctgcag catcgttctg tgttgtctct
      961 gtctgactgt gtttctgtat ttgtctgaga atatgggcca gactgttacc actcccttaa
     1021 gtttgacctt aggtcactgg aaagatgtcg agcggatcgc tcacaaccag tcggtagatg
     1081 tcaagaagag acgttgggtt accttctgct ctgcagaatg gccaaccttt aacgtcggat
     1141 ggccgcgaga cggcaccttt aaccgagacc tcatcaccca ggttaagatc aaggtctttt
     1201 cacctggccc gcatggacac ccagaccagg tcccctacat cgtgacctgg gaagccttgg
     1261 cttttgaccc ccctccctgg gtcaagccct ttgtacaccc taagcctccg cctcctcttc
     1321 ctccatccgc cccgtctctc ccccttgaac ctcctcgttc gaccccgcct cgatcctccc
     1381 tttatccagc cctcactcct tctctaggcg ccaaacctaa acctcaagtt ctttctgaca
     1441 gtggggggcc gctcatcgac ctacttacag aagacccccc gccttatagg gacccaagac
     1501 cacccccttc cgacagggac ggaaatggtg gagaagcgac ccctgcggga gaggcaccgg
     1561 acccctcccc aatggcatct cgcctacgtg ggagacggga gccccctgtg gccgactcca
     1621 ctacctcgca ggcattcccc ctccgcgcag gaggaaacgg acagcttcaa tactggccgt
     1681 tctcctcttc tgacctttac aactggaaaa ataataaccc ttctttttct gaagatccag
     1741 gtaaactgac agctctgatc gagtctgttc tcatcaccca tcagcccacc tgggacgact
     1801 gtcagcagct gttggggact ctgctgaccg gagaagaaaa acaacgggtg ctcttagagg
     1861 ctagaaaggc ggtgcggggc gatgatgggc gccccactca actgcccaat gaagtcgatg
     1921 ccgcttttcc cctcgagaag cttgtcgacg gatccgaatt cgtcgaggga agaaataata
     1981 aagacattaa aatcactttt gtaaactttt taaataaggg aaattgatta aaacaacgcg
     2041 aagttgtaaa atgccatata ctgcatacat tttgtgggaa aaaatgtgca tttacctgct
     2101 caccaaatta tcttattagc tagcagtaga gctctttgtg tgatgggaag atgggataac
     2161 agctgttgtt actgtactgg tttttttttt taatatattt aaaaaaaaga gtgtctccta
     2221 aagtgtaagc gctatcatgc tggactaatg ttttaattta atttaattgt tgtcttggta
     2281 gagatggggt ctcgctattt tgctcacgca gatctcaaac tcctgacgcc caggtacact
     2341 cccatctcgg cctcctaaaa tggtgggatc acagacgtca gccaccgagc ccggtcactt
     2401 ttttgtattc cccacagtat tgatgtatat cttctgcgtt caaaagcaat tttttaaagc
     2461 ctcataacgt ggtaacagaa tactttgcac attacaaaat tcagaacacg gaaacaagaa
     2521 gctcgctttt ttttcccccc tatttcggtt tggcccttta gatttcccct cccccaccgg
     2581 ggcgggactt cccgccgact tctttcaggt tctcagttcg gtccgccaac tgtcgtataa
     2641 aggcgctgcc tcaggccaga ggcctcacaa agcgcgtcga gacagcaagc gaaccggaat
     2701 tgccagctgg ggcgccctct ggtaaggttg ggaagccctg caaagtaaac tggatggctt
     2761 tcttgccgcc aaggatctga tggcgcaggg gatcaagatc gatctgatcc atcgccacca
     2821 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     2881 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     2941 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     3001 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     3061 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     3121 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     3181 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     3241 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     3301 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     3361 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     3421 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     3481 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaaa
     3541 gcggccggct ggatgatcct ccagcgcggg gatctcatgc tggagatccc gcgattagtc
     3601 caatttgtta aagacaggat atcagtggtc caggctctag ttttgactca acaatatcac
     3661 cagctgaagc ctatagagta cgagccatag ataaaataaa agattttatt tagtctccag
     3721 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     3781 ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga
     3841 acagatggaa caggtcgacc tagagaacca tcagatgttt ccagggtgcc ccaaggacct
     3901 gaaatgaccc tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg
     3961 cgcttctgct ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc
     4021 tccgattgac tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc
     4081 cgacttgtgg tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag
     4141 cgggggtctc ttcacatgca gcatgtatca aaattaattt ggtttttttt tcttaagtat
     4201 ttacattaaa tggcccatag tacttaaagt aacatgggct tccttgaaat aaacatggag
     4261 tattcagaat gtgtcataaa tatttctaat tttaagatag tatctccatt ggctatctac
     4321 tttttcttat attttttttt gtcctctgtc ttccattcgt tgttgttgtt gtttgtttgt
     4381 ttgtttgttg gttggttggt taattttttt ttaaagatcc tacactatag ttcaagctag
     4441 actattagct actctgtaac ccagggtgac ctttgaagtc atgggtagcc tgctgtttta
     4501 gccttcccac atctaagatt acaggtatga gctatcattt ttggtatatt gattgattga
     4561 ttgattgatg tgtgtgtgtg tgattgtgtt tgtgtgtgtg attgtgtata tgtgtgtatg
     4621 gttgtgtgtg attgtgtgta tgtatgtttg tgtgtgattg tgtgtgtgtg attgtgcatg
     4681 tgtgtgtgtg tgattgtgtt tatgtgtatg attgtgtgtg tgtgtgtgtg tgtgtgtgtg
     4741 tgtgtgtgtg tgtgtgtgtt gtgtatatat atttatggta gtgagaggca acgctccggc
     4801 tcaggtgtca ggttggtttt tgagacagag tctttcgctt agcttggaat taattcgtaa
     4861 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     4921 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     4981 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     5041 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     5101 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5161 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5221 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5281 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5341 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5401 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5461 caatgctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5521 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5581 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5641 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5701 actagaagga cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5761 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5821 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5881 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5941 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6001 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6061 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6121 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6181 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6241 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6301 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6361 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6421 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6481 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6541 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6601 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6661 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6721 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6781 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6841 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6901 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6961 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7021 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7081 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7141 acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca
     7201 gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg
     7261 agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc
     7321 aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     7381 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7441 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgccaagct agctttgctc
     7501 ttaggagttt cctaatacat cccaaactca aatatataaa gcatttgact tgttctatgc
     7561 cctagggggc ggggggaagc taagccagct ttttttaaca tttaaaatgt taattccatt
     7621 ttaaatgcac agatgttttt atttcataag ggtttcaatg tgcatgaatg ctgcaatatt
     7681 cctgttacca aagctagtat aaataaaaat agataaacgt ggaaattact tagagtttct
     7741 gtcattaacg tttccttcct cagttgacaa cataaatgcg ctgctgagaa gccagtttgc
     7801 atctgtcagg atcaatttcc cattatgcca gtcatattaa ttactagtca attagttgat
     7861 ttttattttt gacatataca tg
//
