LOCUS       sequence-419570-        4479 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of GFP (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4479)
  AUTHORS   Matsuda T, Cepko CL
  TITLE     Electroporation and RNA interference in the rodent retina in vivo 
            and in vitro.
  JOURNAL   Proc Natl Acad Sci U S A. 2004 Jan 6. 101(1):16-22.
  PUBMED    14603031
REFERENCE   2  (bases 1 to 4479)
  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: "Proc Natl 
            Acad Sci U S A. 2004 Jan 6. 101(1):16-22."
COMMENT     Sequence Label: pCMV-GFP
FEATURES             Location/Qualifiers
     source          1..4479
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        129..332
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     283..303
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    355..435
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      437..446
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(443..445,446..448,449..1162)
                     /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(443..445,446..448,449..1159)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(488..509)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(749..768)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1096..1117
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1249..1268)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1314..1369
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1368..1387)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1730..1746)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1730..1746)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1743..1765)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1754..1770
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1778..1784,1785..1802,1803..1808))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1823..1844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1903..2098
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1949..2084
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2104..2238
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2141..2160)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2195..2214
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2306..2323)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2477..3065)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2557..2576)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3236..4027,4028..4096))
                     /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     3859..3878
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4097..4201)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4232..4479,1..128)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
       61 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
      121 ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac
      181 ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc
      241 aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc
      301 gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc
      361 gctaccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc
      421 ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt
      481 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
      541 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
      601 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
      661 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
      721 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
      781 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
      841 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
      901 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
      961 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     1021 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     1081 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     1141 catggacgag ctgtacaagt aaagcggccg cactcctcag gtgcaggctg cctatcagaa
     1201 ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc tttttccctc
     1261 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     1321 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaaggac
     1381 atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga gtttggcaac
     1441 atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc atcagtatat
     1501 gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt gaggttagat
     1561 tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat tttccttaca
     1621 tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag ctgtccctct
     1681 tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg tcatagctgt
     1741 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     1801 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     1861 tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca attagtcagc
     1921 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     1981 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctcggc
     2041 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     2101 gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2161 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2221 tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     2281 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     2341 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     2401 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     2461 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2521 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2581 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     2641 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     2701 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     2761 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     2821 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     2881 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     2941 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     3001 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     3061 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     3121 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     3181 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     3241 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     3301 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     3361 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     3421 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     3481 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     3541 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     3601 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     3661 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     3721 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     3781 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     3841 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     3901 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     3961 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     4021 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     4081 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     4141 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     4201 gcacatttcc ccgaaaagtg ccacctgggt cgacattgat tattgactag taatagtaat
     4261 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
     4321 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
     4381 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
     4441 ggtaaactgc ccacttggca gtacatcaag tgtatcata
//
