LOCUS       sequence-273739-        6516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Fimbrin/Cytoskeleton, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6516)
  AUTHORS   Markwardt ML, Kremers GJ, Kraft CA, Ray K, Cranfill PJ, Wilson KA, 
            Day RN, Wachter RM, Davidson MW, Rizzo MA
  TITLE     An improved cerulean fluorescent protein with enhanced brightness 
            and reduced reversible photoswitching.
  JOURNAL   PLoS One. 2011  . 6(3):e17896.
  PUBMED    21479270
REFERENCE   2  (bases 1 to 6516)
  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: "PLoS One. 
            2011  . 6(3):e17896."
COMMENT     Sequence Label: mCerulean3-Plastin-C-10
FEATURES             Location/Qualifiers
     source          1..6516
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        27..330
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        331..534
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     485..505
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      573..582
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(579..581,582..584,585..1295)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAIHGNVYITADKQKNGIK
                     ANFGLNCNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(624..645)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(885..904)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1232..1253
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     RBS             2657..2665
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     polyA_signal    3342..3463
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3379..3398)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3433..3452
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3470..3925)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3607..3628)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3819..3838
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3952..4056
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(4081..4101)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4125..4335
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        4147..4343
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4194..4329
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4378..5172
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             4378..5172
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4432..4451)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5042..5061
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5360..5379)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5404..5451
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5780..6368
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6269..6288
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
       61 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      121 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      181 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      241 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      301 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      361 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      421 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      481 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta
      541 gtgaaccgtc agatccgcta gcgctaccgg tcgccaccat ggtgagcaag ggcgaggagc
      601 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
      661 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
      721 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgagctggg
      781 gcgtgcagtg cttcgcccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
      841 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
      901 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
      961 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aacgccatcc
     1021 acggcaacgt ctatatcacc gccgacaagc agaagaacgg catcaaggcc aacttcggcc
     1081 tcaactgcaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     1141 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccaagc
     1201 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     1261 ccgggatcac tctcggcatg gacgagctgt acaagtccgg aagcggctcc gggagcggaa
     1321 gcggggaaaa cagtactact accatttctc gggaggagct tgaagaacta caagaggcat
     1381 ttaataaaat agatattgac aatagtgggt atgtcagtga ctatgaactt caagacctgt
     1441 ttaaggaagc aagccttcct ctgcctggct acaaggtgcg cgagattgtg gagaaaattc
     1501 tatcagttgc tgacagcaac aaagatggca aaatcagttt tgaagagttt gtgtcactaa
     1561 tgcaagaatt aaaaagcaaa gatatcagca aaacattccg aaaaataatt aacaagaggg
     1621 aagggattac tgctattgga ggaacttcaa ctatttccag tgagggcaca cagcattctt
     1681 attcagagga agaaaaagtg gcttttgtta actggataaa caaagccctg gagaatgacc
     1741 ctgactgtaa gcatcttata cccatgaatc ccaatgatga tagtcttttc aagtcacttg
     1801 cagatggcat ccttctttgc aaaatgatca acttatctga accagataca attgatgaaa
     1861 gagccatcaa taagaaaaag ctcacgccat tcactatttc tgaaaattta aacctagctc
     1921 tgaattctgc ctcagccatt ggttgtacag tggtcaacat tggtgcatca gatctcaaag
     1981 aaggaaaacc tcacttggtc ttgggacttc tctggcagat catcaaagtt ggcctttttg
     2041 ctgatattga gatttccagg aatgaagctc tgattgcatt gttaaatgaa ggtgaggaac
     2101 tagaggagct gatgaagctt tctcccgagg aattactgct gcgatgggtg aactaccatc
     2161 tgaccaatgc aggatggcat accatcagca acttcagcca agacattaag gactcgagag
     2221 cctattttca tctgcttaat cagattgccc ctaaaggtgg ggaagatgga cctgccattg
     2281 ccattgacct ttcaggaatt aatgagacaa atgacctgaa gcgtgctgga ctcatgcttc
     2341 aagaagcaga taaactgggc tgcaaacagt ttgttactcc tgcagatgtg gtttcaggca
     2401 atcctaaact taatttagct tttgtagcta atttgtttaa cacatacccg tgcctgcaca
     2461 agccgaataa taatgacatc gatatgaatt tactggaagg agagagcaag gaagagagaa
     2521 catttcggaa ctggatgaat tccttgggag tcaacccata cattaatcat ttgtacagtg
     2581 accttgcaga tgctttagtg atctttcagc tctatgagat gatccgagtg ccagtcaact
     2641 ggagccatgt caacaaacct ccttatcctg cccttggagg gaacatgaag aagattgaaa
     2701 actgtaacta tgcagtggaa cttgggaaga acaaggccaa attctccttg gttggcattg
     2761 ctgggcagga cctaaatgaa gggaattcaa cacttaccct ggcattggta tggcagctga
     2821 tgagaaggta cacattgaat gtgttatcgg atcttggaga gggtgaaaaa gtaaatgatg
     2881 aaattataat taaatgggtc aatcagactc ttaaaagtgc aaacaaaaag acttctattt
     2941 ccagcttcaa ggataaatct ataagcacaa gtttacctgt cctagattta atagatgcca
     3001 ttgcaccaaa tgcagttcgt caagaaatga tcaggagaga aaacttatct gatgaggaca
     3061 agctgaacaa tgctaaatac gccatttcag ttgctcgaaa gatcggtgcc cggatatatg
     3121 cattacctga tgacctcgta gaagtgaaac caaagatggt tatgacggtg tttgcatgct
     3181 taatgggaaa aggactgaac agaataaaat aaggatccac cggatctaga taactgatca
     3241 taatcagcca taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc
     3301 ccctgaacct gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt
     3361 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3421 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc
     3481 gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa
     3541 taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt
     3601 gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg
     3661 cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt
     3721 ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga
     3781 gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg
     3841 ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg
     3901 cttaatgcgc cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     3961 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     4021 tgataaatgc ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt
     4081 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     4141 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     4201 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     4261 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     4321 ttggaggcct aggcttttgc aaagatcgat caagagacag gatgaggatc gtttcgcatg
     4381 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     4441 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     4501 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa
     4561 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     4621 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     4681 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     4741 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     4801 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     4861 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgagcat gcccgacggc
     4921 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     4981 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     5041 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     5101 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     5161 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     5221 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     5281 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     5341 accctagggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     5401 tgacggcaat aaaaagacag aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg
     5461 gggttcggtc ccagggctgg cactctgtcg ataccccacc gagaccccat tggggccaat
     5521 acgcccgcgt ttcttccttt tccccacccc accccccaag ttcgggtgaa ggcccagggc
     5581 tcgcagccaa cgtcggggcg gcaggccctg ccatagcctc aggttactca tatatacttt
     5641 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     5701 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     5761 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     5821 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     5881 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     5941 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     6001 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     6061 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     6121 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     6181 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     6241 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     6301 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     6361 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     6421 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     6481 attagttatt aatagtaatc aattacgggg tcatta
//
