LOCUS       sequence-255087-        3884 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP reporter vector with restriction sites to clone in a promoter 
            of interest, derived from pGL3 basic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3884)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 3884)
  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: pGL3 basic EGFP
FEATURES             Location/Qualifiers
     source          1..3884
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(87..89,90..92,93..806)
                     /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(87..89,90..92,93..803)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(132..153)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(393..412)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     740..761
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    847..968
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(858..877)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     912..931
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1216..1233)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1387..1975)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1467..1486)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2146..2937,2938..3006))
                     /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     2769..2788
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3007..3111)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3138..3593
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3225..3244)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3435..3456
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3724..3772
                     /note="synthetic polyadenylation signal"
     misc_feature    3786..3877
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     3826..3845
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctaa gtaagcttgg
       61 cattccggta ctgttggtaa accaccatgg tgagcaaggg cgaggagctg ttcaccgggg
      121 tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg
      181 gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg
      241 gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct
      301 tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag
      361 gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg
      421 aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca
      481 aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct
      541 atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca
      601 tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg
      661 gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc
      721 ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc
      781 tcggcatgga cgagctgtac aagtaattct agagtcgggg cggccggccg cttcgagcag
      841 acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag tgaaaaaaat
      901 gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata agctgcaata
      961 aacaagttaa caacaacaat tgcattcatt ttatgtttca ggttcagggg gaggtgtggg
     1021 aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag gatccgtcga
     1081 ccgatgccct tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact
     1141 atcgtcgccg cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca
     1201 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     1261 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     1321 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     1381 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     1441 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     1501 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     1561 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     1621 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     1681 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     1741 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     1801 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     1861 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     1921 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     1981 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     2041 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     2101 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     2161 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     2221 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     2281 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     2341 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     2401 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     2461 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     2521 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     2581 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     2641 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     2701 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     2761 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     2821 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     2881 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     2941 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     3001 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     3061 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     3121 ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     3181 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     3241 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     3301 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     3361 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     3421 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     3481 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     3541 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttgccattc
     3601 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     3661 ccagcccaag ctaccatgat aagtaagtaa tattaaggta cgggaggtac ttggagcggc
     3721 cgcaataaaa tatctttatt ttcattacat ctgtgtgttg gttttttgtg tgaatcgata
     3781 gtactaacat acgctctcca tcaaaacaaa acgaaacaaa acaaactagc aaaataggct
     3841 gtccccagtg caagtgcagg tgccagaaca tttctctatc gata
//
