LOCUS       sequence-248430-        2649 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Minimal backbone plasmid for mammalian eGFP expression from SV40 
            promoter with zeocin/bleocin bacterial resistance.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2649)
  TITLE     Piggybac transposon plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2649)
  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: 
            "Unpublished"
COMMENT     Sequence Label: SV40-eGFP-Z1
FEATURES             Location/Qualifiers
     source          1..2649
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..216
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(48..68)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        66..363
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      233..368
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      391..400
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(397..399,400..402,403..1116)
                     /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(397..399,400..402,403..1113)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(442..463)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(703..722)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1050..1071
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1166..1287
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1177..1196)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1231..1250
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1446..1820)
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     promoter        complement(1830..1848)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1840..1886)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(1959..2547)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2039..2058)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcagaagc tttactaact agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
       61 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      121 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
      181 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
      241 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      301 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
      361 tggaggccta gggggtccgt acgaccggtc gccaccatgg tgagcaaggg cgaggagctg
      421 ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg ccacaagttc
      481 agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct gaagttcatc
      541 tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct gacctacggc
      601 gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt caagtccgcc
      661 atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg caactacaag
      721 acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga gctgaagggc
      781 atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa ctacaacagc
      841 cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa cttcaagatc
      901 cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca gaacaccccc
      961 atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca gtccgccctg
     1021 agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc
     1081 gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc tagatgtaca
     1141 catatggcta gctggccaga catgataaga tacattgatg agtttggaca aaccacaact
     1201 agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta
     1261 accattataa gctgcaataa acaagttaac aacaacaatt gcattcattt tatgtttcag
     1321 gttcaggggg aggtgtggga ggttttttaa agcaagtaaa acctctacaa atgtggtatg
     1381 gaattctgga ctcacaaaga aaaaacgccc ggtgtgcaag accgagcgtt ctgaacaatt
     1441 tatcctcagt cctgctcctc tgccacaaag tgcacgcagt tgccggccgg gtcgcgcagg
     1501 gcgaactccc gcccccacgg ctgctcgccg atctcggtca tggccggccc ggaggcgtcc
     1561 cggaagttcg tggacacgac ctccgaccac tcggcgtaca gctcgtccag gccgcgcacc
     1621 cacacccagg ccagggtgtt gtccggcacc acctggtcct ggaccgcgct gatgaacagg
     1681 gtcacgtcgt cccggaccac accggcgaag tcgtcctcca cgaagtcccg ggagaacccg
     1741 agccggtcgg tccagaactc gaccgctccg gcgacgtcgc gcgcggtgag caccggaacg
     1801 gcactggtca acttggccat gatggccctc ctatagtgag tcgtattata ctatgccgat
     1861 atactatgcc gatgattaat tgtcaaggat cctaagaaca tgtgagcaaa aggccagcaa
     1921 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     1981 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2041 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2101 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2161 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2221 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2281 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2341 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     2401 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     2461 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     2521 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     2581 gctcagtgaa aactcacgtt aagggatttt ggtcatggct agttaattaa catttaaatc
     2641 agcggccgc
//
