LOCUS       sequence-261776-        2869 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhanced green fluorescent protein expression for mammalian cells 
            under EF1a-HTLV promoter in minimal backbone plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2869)
  TITLE     Constitutive promoter minimal backbone plasmids for optimized 
            expression in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2869)
  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            "Unpublished"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: EF1-alpha-eGFP-Z1
FEATURES             Location/Qualifiers
     source          1..2869
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        54..265
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             278..546
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human 
                     T-cell leukemia virus (HTLV) type 1"
     regulatory      611..620
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(617..619,620..622,623..1336)
                     /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(617..619,620..622,623..1333)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(662..683)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(923..942)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1270..1291
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1386..1507
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1397..1416)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1451..1470
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1666..2040)
                     /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(2050..2068)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2060..2106)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(2179..2767)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2259..2278)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcagaagc tttactaact agtggatctg cgatcgctcc ggtgcccgtc agtgggcaga
       61 gcgcacatcg cccacagtcc ccgagaagtt ggggggaggg gtcggcaatt gaacgggtgc
      121 ctagagaagg tggcgcgggg taaactggga aagtgatgtc gtgtactggc tccgcctttt
      181 tcccgagggt gggggagaac cgtatataag tgcagtagtc gccgtgaacg ttctttttcg
      241 caacgggttt gccgccagaa cacagctgaa gcttcgaggg gctcgcatct ctccttcacg
      301 cgcccgccgc cctacctgag gccgccatcc acgccggttg agtcgcgttc tgccgcctcc
      361 cgcctgtggt gcctcctgaa ctgcgtccgc cgtctaggta agtttaaagc tcaggtcgag
      421 accgggcctt tgtccggcgc tcccttggag cctacctaga ctcagccggc tctccacgct
      481 ttgcctgacc ctgcttgctc aactctacgt ctttgtttcg ttttctgttc tgcgccgtta
      541 cagatccaag ctgtgaccgg cgcctacaat tctgcagtcg acggtaccgc gggcccggga
      601 tccaccggtc gccaccatgg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
      661 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
      721 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
      781 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
      841 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
      901 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
      961 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     1021 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     1081 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     1141 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     1201 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     1261 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     1321 cgagctgtac aagtaaagcg gccgcgactc tagatgtaca catatggcta gctggccaga
     1381 catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt gaaaaaaatg
     1441 ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa gctgcaataa
     1501 acaagttaac aacaacaatt gcattcattt tatgtttcag gttcaggggg aggtgtggga
     1561 ggttttttaa agcaagtaaa acctctacaa atgtggtatg gaattctgga ctcacaaaga
     1621 aaaaacgccc ggtgtgcaag accgagcgtt ctgaacaatt tatcctcagt cctgctcctc
     1681 tgccacaaag tgcacgcagt tgccggccgg gtcgcgcagg gcgaactccc gcccccacgg
     1741 ctgctcgccg atctcggtca tggccggccc ggaggcgtcc cggaagttcg tggacacgac
     1801 ctccgaccac tcggcgtaca gctcgtccag gccgcgcacc cacacccagg ccagggtgtt
     1861 gtccggcacc acctggtcct ggaccgcgct gatgaacagg gtcacgtcgt cccggaccac
     1921 accggcgaag tcgtcctcca cgaagtcccg ggagaacccg agccggtcgg tccagaactc
     1981 gaccgctccg gcgacgtcgc gcgcggtgag caccggaacg gcactggtca acttggccat
     2041 gatggccctc ctatagtgag tcgtattata ctatgccgat atactatgcc gatgattaat
     2101 tgtcaaggat cctaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     2161 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     2221 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     2281 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     2341 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     2401 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     2461 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     2521 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     2581 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     2641 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     2701 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     2761 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgaa aactcacgtt
     2821 aagggatttt ggtcatggct agttaattaa catttaaatc agcggccgc
//
