LOCUS       sequence-217334-        4509 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of CFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4509)
  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 4509)
  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: pCAG-CFP
FEATURES             Location/Qualifiers
     source          1..4509
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     primer_bind     38..60
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     122..141
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      207..216
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(213..215,216..218,219..932)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(258..279)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(519..538)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     866..887
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1019..1038)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1084..1139
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1138..1157)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1500..1516)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1500..1516)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1513..1535)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1524..1540
                     /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(1548..1554,1555..1572,1573..1578))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1593..1614
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1673..1868
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1719..1854
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1874..2008
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1911..1930)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1965..1984
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2076..2093)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2247..2835)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2327..2346)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3006..3797,3798..3866))
                     /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     3629..3648
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3867..3971)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4001..4380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
       61 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
      121 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctg cagtcgacgg
      181 taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
      241 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
      301 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
      361 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tggggcgtgc
      421 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
      481 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
      541 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
      601 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac atcagccaca
      661 acgtctatat caccgccgac aagcagaaga acggcatcaa ggccaacttc aagatccgcc
      721 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
      781 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
      841 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
      901 tcactctcgg catggacgag ctgtacaagt aaagcggccg cactcctcag gtgcaggctg
      961 cctatcagaa ggtggtggct ggtgtggcca atgccctggc tcacaaatac cactgagatc
     1021 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     1081 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     1141 tcggaaggac atatgggagg gcaaatcatt taaaacatca gaatgagtat ttggtttaga
     1201 gtttggcaac atatgcccat atgctggctg ccatgaacaa aggttggcta taaagaggtc
     1261 atcagtatat gaaacagccc cctgctgtcc attccttatt ccatagaaaa gccttgactt
     1321 gaggttagat tttttttata ttttgttttg tgttattttt ttctttaaca tccctaaaat
     1381 tttccttaca tgttttacta gccagatttt tcctcctctc ctgactactc ccagtcatag
     1441 ctgtccctct tctcttatgg agatccctcg acctgcagcc caagcttggc gtaatcatgg
     1501 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     1561 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     1621 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagcggat ccgcatctca
     1681 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     1741 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     1801 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     1861 tttgcaaaaa gctaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     1921 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     1981 catcaatgta tcttatcatg tctggatccg ctgcattaat gaatcggcca acgcgcgggg
     2041 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2101 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2161 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2221 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2281 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2341 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2401 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2461 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2521 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2581 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2641 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2701 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2761 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2821 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2881 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2941 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3001 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3061 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3121 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3181 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3241 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3301 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3361 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3421 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3481 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3541 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3601 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3661 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3721 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3781 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3841 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3901 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3961 ggggttccgc gcacatttcc ccgaaaagtg ccacctggtc gacattgatt attgactagt
     4021 tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt
     4081 acataactta cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg
     4141 tcaataatga cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg
     4201 gtggagtatt tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt
     4261 acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg
     4321 accttatggg actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg
     4381 gtcgaggtga gccccacgtt ctgcttcact ctccccatct cccccccctc cccaccccca
     4441 attttgtatt tatttatttt ttaattattt tgtgcagcga tgggggcggg gggggggggg
     4501 gcgcgcgcc
//
