LOCUS       sequence-406946-        6127 bp    DNA     circular SYN 14-APR-2025
DEFINITION  mammalian expression of mSECFP, a monomeric super enhanced cyan 
            fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6127)
  AUTHORS   Matsuda T, Miyawaki A, Nagai T
  TITLE     Direct measurement of protein dynamics inside cells using a 
            rationally designed photoconvertible protein.
  JOURNAL   Nat Methods. 2008 Apr;5(4):339-45. doi: 10.1038/nmeth.1193. Epub 
            2008 Mar 16.
  PUBMED    18345008
REFERENCE   2  (bases 1 to 6127)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/nmeth.1193"; journalName: "Nat Methods"; date: "2008-04"; 
            volume: "5"; issue: "4"; pages: "339-45"
COMMENT     Sequence Label: mSECFP/pcDNA3
FEATURES             Location/Qualifiers
     source          1..6127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(666..668,669..671,672..1385)
                     /codon_start=1
                     /product="monomeric super enhanced variant of CFP (Matsuda 
                     et al., 2008)"
                     /label=mseCFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(711..732)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(972..991)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1319..1340
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(1446..1464)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1447..1464)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1484..1501)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1490..1714
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1760..2188
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1847..1866)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2057..2078
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2197..2217)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2202..2531
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2382..2517
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2444..2463
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2598..3392
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2652..2671)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3262..3281
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3566..3687
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3603..3622)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3657..3676
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3736..3752)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3736..3752)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3749..3771)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3760..3776
                     /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(3784..3790,3791..3808,3809..3814))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3829..3850
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3967..3984)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4138..4726)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4218..4237)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4897..5688,5689..5757))
                     /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     5520..5539
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5758..5862)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5937..5956)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      721 tggacggcga cgtaaacggc cacaggttca gcgtgtccgg cgagggcgag ggcgatgcca
      781 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      841 ccaccctcgt gaccaccctg acctggggcg tgcagtgctt cgcccgctac cccgaccaca
      901 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgtacca
      961 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     1021 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     1081 ggcacaagct ggagtacaac tacatcagcc acaacgtcta tatcaccgcc gacaagcaga
     1141 agaacggcat caaggcccac ttcaagatcc gccacaacat cgaggacggc ggcgtgcagc
     1201 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1261 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     1321 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1381 agtaagaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc
     1441 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     1501 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1561 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1621 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1681 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     1741 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     1801 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     1861 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     1921 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     1981 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2041 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2101 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2161 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2221 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2281 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2341 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2401 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2461 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2521 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     2581 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     2641 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     2701 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     2761 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     2821 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     2881 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     2941 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     3001 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     3061 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     3121 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     3181 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     3241 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     3301 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     3361 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     3421 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     3481 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     3541 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     3601 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     3661 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     3721 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     3781 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     3841 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     3901 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     3961 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     4021 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     4081 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     4141 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     4201 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     4261 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     4321 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     4381 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     4441 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     4501 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     4561 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     4621 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     4681 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     4741 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     4801 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     4861 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     4921 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     4981 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     5041 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     5101 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     5161 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     5221 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     5281 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     5341 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     5401 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     5461 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     5521 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     5581 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     5641 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     5701 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     5761 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     5821 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     5881 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     5941 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     6001 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     6061 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     6121 acgcgtt
//
