LOCUS       sequence-269408-        2946 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhanced Green fluorescent protein expression for mammalian cells 
            under constitutive CMV Promoter in minimal backbone plasmid..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2946)
  TITLE     Constitutive promoter minimal backbone plasmids for optimized 
            expression in mammalian cells
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2946)
  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            "Unpublished"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: CMV-eGFP-Z1
FEATURES             Location/Qualifiers
     source          1..2946
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        82..385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..589
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     540..560
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    612..692
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      694..703
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(700..702,703..705,706..1419)
                     /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(700..702,703..705,706..1416)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(745..766)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1006..1025)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1353..1374
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1469..1590
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1480..1499)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1534..1553
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1749..2123)
                     /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(2133..2151)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(2143..2189)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     rep_origin      complement(2262..2850)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2342..2361)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccgcctgc agaagcttta ctaactagta atagtaatca attacggggt cattagttca
       61 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
      121 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
      181 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
      241 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
      301 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
      361 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
      421 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
      481 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
      541 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa
      601 ccgtcagatc cgctagcgct accggactca gatctcgagc tcaagcttcg aattctgcag
      661 tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
      721 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
      781 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
      841 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
      901 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
      961 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     1021 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     1081 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     1141 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     1201 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     1261 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     1321 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     1381 gccgggatca ctctcggcat ggacgagctg tacaagtaaa gcggccgcga ctctagatgt
     1441 acacatatgg ctagctggcc agacatgata agatacattg atgagtttgg acaaaccaca
     1501 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1561 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     1621 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     1681 atggaattct ggactcacaa agaaaaaacg cccggtgtgc aagaccgagc gttctgaaca
     1741 atttatcctc agtcctgctc ctctgccaca aagtgcacgc agttgccggc cgggtcgcgc
     1801 agggcgaact cccgccccca cggctgctcg ccgatctcgg tcatggccgg cccggaggcg
     1861 tcccggaagt tcgtggacac gacctccgac cactcggcgt acagctcgtc caggccgcgc
     1921 acccacaccc aggccagggt gttgtccggc accacctggt cctggaccgc gctgatgaac
     1981 agggtcacgt cgtcccggac cacaccggcg aagtcgtcct ccacgaagtc ccgggagaac
     2041 ccgagccggt cggtccagaa ctcgaccgct ccggcgacgt cgcgcgcggt gagcaccgga
     2101 acggcactgg tcaacttggc catgatggcc ctcctatagt gagtcgtatt atactatgcc
     2161 gatatactat gccgatgatt aattgtcaag gatcctaaga acatgtgagc aaaaggccag
     2221 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     2281 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     2341 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     2401 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     2461 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     2521 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     2581 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     2641 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     2701 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     2761 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     2821 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     2881 gacgctcagt gaaaactcac gttaagggat tttggtcatg gctagttaat taacatttaa
     2941 atcagc
//
