LOCUS       sequence-255091-        4077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP reporter vector with restriction sites to clone in an enhancer 
            of interest, derived from pGL3 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4077)
  TITLE     Emory Custom Cloning Core Plasmids - Oskar Laur
REFERENCE   2  (bases 1 to 4077)
  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 promoter  EGFP
FEATURES             Location/Qualifiers
     source          1..4077
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..244
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      95..230
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      274..283
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(280..282,283..285,286..999)
                     /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(280..282,283..285,286..996)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(325..346)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(586..605)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     933..954
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1040..1161
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1051..1070)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1105..1124
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1409..1426)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1580..2168)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1660..1679)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2339..3130,3131..3199))
                     /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     2962..2981
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3200..3304)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3331..3786
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3418..3437)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3628..3649
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    3917..3965
                     /note="synthetic polyadenylation signal"
     misc_feature    3979..4070
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     4019..4038
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
ORIGIN
        1 ggtaccgagc tcttacgcgt gctagcccgg gctcgagatc tgcgatctgc atctcaatta
       61 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
      121 cgcccattct ccgccccatc gctgactaat tttttttatt tatgcagagg ccgaggccgc
      181 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
      241 caaaaagctt ggcattccgg tactgttggt aaagccacca tggtgagcaa gggcgaggag
      301 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      361 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      421 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
      481 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      541 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
      601 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
      661 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
      721 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
      781 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
      841 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
      901 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
      961 gccgggatca ctctcggcat ggacgagctg tacaagtaat tctagagtcg gggcggccgg
     1021 ccgcttcgag cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     1081 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     1141 ataagctgca ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag
     1201 ggggaggtgt gggaggtttt ttaaagcaag taaaacctct acaaatgtgg taaaatcgat
     1261 aaggatccgt cgaccgatgc ccttgagagc cttcaaccca gtcagctcct tccggtgggc
     1321 gcggggcatg actatcgtcg ccgcacttat gactgtcttc tttatcatgc aactcgtagg
     1381 acaggtgccg gcagcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1441 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1501 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1561 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1621 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1681 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1741 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1801 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1861 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1921 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1981 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     2041 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2101 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2161 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2221 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2281 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2341 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2401 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2461 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2521 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2581 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2641 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2701 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2761 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2821 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2881 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2941 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3001 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3061 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3121 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3181 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3241 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3301 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     3361 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3421 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3481 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3541 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3601 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3661 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3721 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3781 caatttgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3841 cttcgctatt acgccagccc aagctaccat gataagtaag taatattaag gtacgggagg
     3901 tacttggagc ggccgcaata aaatatcttt attttcatta catctgtgtg ttggtttttt
     3961 gtgtgaatcg atagtactaa catacgctct ccatcaaaac aaaacgaaac aaaacaaact
     4021 agcaaaatag gctgtcccca gtgcaagtgc aggtgccaga acatttctct atcgata
//
