LOCUS       sequence-325843-        5857 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Introduce constitutive (SFFV promoter) GFP cDNA into (ES) cells by 
            recombination-mediated cassette exchange.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5857)
  TITLE     Karreth lab plasmids
REFERENCE   2  (bases 1 to 5857)
  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"
COMMENT     Sequence Label: cSFFV-GFP
FEATURES             Location/Qualifiers
     source          1..5857
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..547
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(83..106)
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
     primer_bind     480..499
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     protein_bind    593..640
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    join(673..738,739..776)
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     polyA_signal    945..1079
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    1199..1333
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        1349..1755
                     /label=SFFV promoter
                     /note="spleen focus-forming virus long terminal repeat 
                     (LTR) promoter"
     CDS             join(1762..1764,1765..1767,1768..2481)
                     /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(1762..1764,1765..1767,1768..2478)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1807..1828)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2068..2087)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2415..2436
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     intron          2524..3096
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     rep_origin      complement(3424..4012)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3504..3523)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4183..4974,4975..5043))
                     /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     4806..4825
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5044..5148)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5174..5629)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5311..5332)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5523..5542
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5756..5778
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     5770..5787
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5771..5787
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5794..5813
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5794..5812
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     join(5845..5857,1..4)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(5845..5857,1..4)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
ORIGIN
        1 gatcccccgg gctgcaggaa ttaattcgtc gacctcgaaa ttctaccggg taggggaggc
       61 gcttttccca aggcagtctg gagcatgcgc tttagcagcc ccgctgggca cttggcgcta
      121 cacaagtggc ctctggcctc gcacacattc cacatccacc ggtaggcgcc aaccggctcc
      181 gttctttggt ggccccttcg cgccaccttc tactcctccc ctagtcagga agttcccccc
      241 cgccccgcag ctcgcgtcgt gcaggacgtg acaaatggaa gtagcacgtc tcactagtct
      301 cgtgcagatg gacagcaccg ctgagcaatg gaagcgggta ggcctttggg gcagcggcca
      361 atagcagctt tgctccttcg ctttctgggc tcagaggctg ggaaggggtg ggtccggggg
      421 cgggctcagg ggcgggctca ggggcggggc gggcgcccga aggtcctccg gaggcccggc
      481 attctgcacg cttcaaaagc gcacgtctgc cgcgctgttc tcctcttcct catctccggg
      541 cctttcgacc tgcatccatc tagatctcga tcgagcagct gaagcttatg gagaagttac
      601 tattccgaag ttcctattct ctagaaagta taggaacttc gatccccctc gatcccccgg
      661 gctgcagatc tgtagggcgc agtagtccag ggtttccttg atgatgtcat acttatcctg
      721 tccctttttt ttccacagct cgcggttgag gacaaactct tcgcggtctt tccagtgggg
      781 atcgacggta tcgataagct tgcagatctg cgactctaga ggatctgcga ctctagagga
      841 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
      901 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
      961 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     1021 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatct
     1081 gcgactctag aggatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1141 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1201 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1261 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1321 tcatgtctgg atcgagttta ccatgcatgt aacgccattt tgcaaggcat ggaaaaatac
     1381 caaaccaaga atagagaagt tcagatcaag ggcgggtaca tgaaaatagc taacgttggg
     1441 ccaaacagga tatctgcggt gagcagtttc ggccccggcc cggggccaag aacagatggt
     1501 caccgcagtt tcggccccgg cccgaggcca agaacagatg gtccccagat atggcccaac
     1561 cctcagcagt ttcttaagac ccatcagatg tttccaggct cccccaagga cctgaaatga
     1621 ccctgcgcct tatttgaatt aaccaatcag cctgcttctc gcttctgttc gcgcgcttct
     1681 gcttcccgag ctctataaaa gagctcacaa cccctcactc ggcgcgccag tcctccgaca
     1741 gactgagtcg gccggtccac catggtgagc aagggcgagg agctgttcac cggggtggtg
     1801 cccatcctgg tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag
     1861 ggcgagggcg atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag
     1921 ctgcccgtgc cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc
     1981 cgctaccccg accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac
     2041 gtccaggagc gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg
     2101 aagttcgagg gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag
     2161 gacggcaaca tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc
     2221 atggccgaca agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag
     2281 gacggcagcg tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc
     2341 gtgctgctgc ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac
     2401 gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc
     2461 atggacgagc tgtacaagta actcgagttc atgaattcgc agtcgagcgt tgagaacttc
     2521 agggtgagtt tggggaccct tgattgttct ttctttttcg ctattgtaaa attcatgtta
     2581 tatggagggg gcaaagtttt cagggtgttg tttagaatgg gaagatgtcc cttgtatcac
     2641 catggaccct catgataatt ttgtttcttt cactttctac tctgttgaca accattgtct
     2701 cctcttattt tcttttcatt ttctgtaact ttttcgttaa actttagctt gcatttgtaa
     2761 cgaattttta aattcacttt tgtttatttg tcagattgta agtactttct ctaatcactt
     2821 ttttttcaag gcaatcaggg tatattatat tgtacttcag cacagtttta gagaacaatt
     2881 gttataatta aatgataagg tagaatattt ctgcatataa attctggctg gcgtggaaat
     2941 attcttattg gtagaaacaa ctacaccctg gtcatcatcc tgcctttctc tttatggtta
     3001 caatgatata cactgtttga gatgaggata aaatactctg agtccaaacc gggcccctct
     3061 gctaaccatg ttcatgcctt cttctctttc ctacagctcc tgggcaacgt gctggttgtt
     3121 gtgctgtctc atcattttgg caaaggattc actcctcagg tgcaggctgc ctatcagaag
     3181 gtggtggctg gtgtggccaa tgccctggct cacaaatacc actgagatcg ttttccctct
     3241 gccaaaaatt atggggacat catgaagccc cttgagcatc tgacttctgg ctaataaagg
     3301 aaatttattt tcattgcaat agtgtgttgg aatttctcga tcgctagcat cctaggcgcg
     3361 ccacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     3421 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     3481 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     3541 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     3601 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     3661 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     3721 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     3781 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     3841 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     3901 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     3961 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4021 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4081 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     4141 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     4201 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     4261 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     4321 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     4381 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     4441 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     4501 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     4561 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     4621 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     4681 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     4741 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     4801 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     4861 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     4921 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     4981 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     5041 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     5101 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     5161 aaaagtgcca cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt
     5221 taaatcagct cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa
     5281 gaatagaccg agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag
     5341 aacgtggact ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt
     5401 gaaccatcac cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac
     5461 cctaaaggga gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag
     5521 gaagggaaga aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg
     5581 cgcgtaacca ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca
     5641 ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag
     5701 ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag
     5761 tcacgacgtt gtaaaacgac ggccagtgaa ttgtaatacg actcactata gggcgaattg
     5821 gagctccacc gcggtggcgg ccgctctaga actagtg
//
