LOCUS       sequence-334928-        3930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GFP under the control of EF1a short promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3930)
  AUTHORS   Ream MW, Randolph LN, Jiang Y, Chang Y, Bao X, Lian XL
  TITLE     Direct programming of human pluripotent stem cells into endothelial 
            progenitors with SOX17 and FGF2.
  JOURNAL   Stem Cell Reports. 2024 Mar 5:S2213-6711(24)00044-4. doi: 
            10.1016/j.stemcr.2024.02.006.
  PUBMED    38518781
REFERENCE   2  (bases 1 to 3930)
  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: "Stem Cell 
            Reports. 2024 Mar 5:S2213-6711(24)00044-4. doi: 
            10.1016/j.stemcr.2024.02.006."
COMMENT     Sequence Label: AAV-EFS-GFP
FEATURES             Location/Qualifiers
     source          1..3930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        175..386
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     CDS             join(411..413,414..416,417..1130)
                     /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(411..413,414..416,417..1127)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(456..477)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(717..736)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1064..1085
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1137..1185
                     /note="synthetic polyadenylation signal"
     repeat_region   1193..1333
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1193..1322
                     /label=AAV2 ITR
     rep_origin      1408..1863
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1495..1514)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1705..1726
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1880..1899)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1999..2021
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(2059..2077)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2145..2249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2250..2318,2319..3110)
                     /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     complement(2468..2487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3281..3869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3770..3789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggcctcta gaaaggatct gcgatcgctc cggtgcccgt cagtgggcag
      181 agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat tgaacgggtg
      241 cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt
      301 ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac gttctttttc
      361 gcaacgggtt tgccgccaga acacagctga agcttggtat cgataccggt atggtgagca
      421 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      481 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      541 ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
      601 ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact
      661 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
      721 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
      781 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
      841 acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg
      901 tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc
      961 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca
     1021 cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1081 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa gaattcaata
     1141 aaagatcttt attttcatta gatctgtgtg ttggtttttt gtgtgcggcc gcaggaaccc
     1201 ctagtgatgg agttggccac tccctctctg cgcgctcgct cgctcactga ggccgggcga
     1261 ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct cagtgagcga gcgagcgcgc
     1321 agctgcctgc aggggcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca
     1381 caccgcatac gtcaaagcaa ccatagtacg cgccctgtag cggcgcatta agcgcggcgg
     1441 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccttagcg cccgctcctt
     1501 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1561 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1621 atttgggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1681 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaact
     1741 ctatctcggg ctattctttt gatttataag ggattttgcc gatttcggtc tattggttaa
     1801 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgtttacaa
     1861 ttttatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagccccgac
     1921 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     1981 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     2041 aacgcgcgag acgaaagggc ctcgtgatac gcctattttt ataggttaat gtcatgataa
     2101 taatggtttc ttagacgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     2161 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     2221 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     2281 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     2341 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     2401 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     2461 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     2521 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     2581 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     2641 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     2701 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     2761 taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac
     2821 tattaactgg cgaactactt actctagctt cccggcaaca attaatagac tggatggagg
     2881 cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg tttattgctg
     2941 ataaatctgg agccggtgag cgtggaagcc gcggtatcat tgcagcactg gggccagatg
     3001 gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact atggatgaac
     3061 gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa ctgtcagacc
     3121 aagtttactc atatatactt tagattgatt taaaacttca tttttaattt aaaaggatct
     3181 aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc
     3241 actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct ttttttctgc
     3301 gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt tgtttgccgg
     3361 atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg cagataccaa
     3421 atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct gtagcaccgc
     3481 ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc gataagtcgt
     3541 gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcgggctgaa
     3601 cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa ctgagatacc
     3661 tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc
     3721 cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg ggaaacgcct
     3781 ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga tttttgtgat
     3841 gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt ttacggttcc
     3901 tggccttttg ctggcctttt gctcacatgt
//
