LOCUS       sequence-468016-        6309 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Membrane/Vesicles, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6309)
  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 6309)
  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-C-Src-7
FEATURES             Location/Qualifiers
     source          1..6309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             575..616
                     /codon_start=1
                     /product="N-myristoylation signal from Src kinase (Pellman 
                     et al., 1985; Kaplan et al., 1988)"
                     /label=myr
                     /translation="MGSSKSKPKDPSQR"
     regulatory      2189..2198
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2195..2197,2198..2200,2201..2914)
                     /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(2240..2261)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2501..2520)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2848..2869
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3037..3158
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3074..3093)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3128..3147
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3165..3620)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3302..3323)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3514..3533
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3647..3751
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3753..4110
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3753..3944
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3776..3796)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3961..4096
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4145..4939
                     /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             4145..4939
                     /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(4199..4218)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4809..4828
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5127..5146)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5171..5218
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5547..6135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6036..6055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      541 ccggactcag atctcgagct caagcttcga attcatgggg agcagcaaga gcaagcccaa
      601 ggaccccagc cagcgccggc gcagcctgga gccacccgac agcacccacc acgggggatt
      661 cccagcctcg cagaccccca acaagacagc agcccccgac acgcaccgca cccccagccg
      721 ctcctttggg accgtggcca ccgagcccaa gctcttcggg ggcttcaaca cttctgacac
      781 cgtcacgtcg ccgcagcgtg ccggggcact ggctggcggc gtcaccactt tcgtggctct
      841 ctatgactac gagtcccgga ctgaaacgga cttgtccttc aagaaaggag aacgcctgca
      901 gattgtcaac aacacggaag gtgactggtg gctggctcat tccctcacta caggacagac
      961 gggctacatc cccagtaact atgtcgcgcc ctcagactcc atccaggctg aagagtggta
     1021 ctttgggaag atcactcgtc gggagtccga gcggctgctg ctcaaccccg aaaacccccg
     1081 gggaaccttc ttggtccggg agagcgagac gacaaaaggt gcctattgcc tctccgtttc
     1141 tgactttgac aacgccaagg ggctcaatgt gaagcactac aagatccgca agctggacag
     1201 cggcggcttc tacatcacct cacgcacaca gttcagcagc ctgcagcagc tggtggccta
     1261 ctactccaaa catgctgatg gcttgtgcca ccgcctgacc aacgtctgcc ccacgtccaa
     1321 gccccagacc cagggactcg ccaaggacgc gtgggaaatc ccccgggagt cgctgcggct
     1381 ggaggtgaag ctggggcagg gctgctttgg agaggtctgg atggggacct ggaacggcac
     1441 caccagagtg gccataaaga ctctgaagcc cggcaccatg tccccggagg ccttcctgca
     1501 ggaagcccaa gtgatgaaga agctccggca tgagaagctg gttcagctgt acgcagtggt
     1561 gtcggaagag cccatctaca tcgtcactga gtacatgagc aaggggagcc tcctggattt
     1621 cctgaaggga gagatgggca agtacctgcg gctgccacag ctcgtcgata tggctgctca
     1681 gattgcatcc ggcatggcct atgtggagag gatgaactac gtgcaccgag acctgcgggc
     1741 ggccaacatc ctggtggggg agaacctggt gtgcaaggtg gctgactttg ggctggcacg
     1801 cctcatcgag gacaacgagt acacagcacg gcaaggtgcc aagttcccca tcaagtggac
     1861 agcccccgag gcagccctct atggccggtt caccatcaag tcggatgtct ggtccttcgg
     1921 catcctgctg actgagctga ccaccaaggg ccgggtgcca tacccaggga tggtcaacag
     1981 ggaggtgctg gaccaggtgg agaggggcta ccgcatgccc tgcccgcccg agtgccccga
     2041 gtcgctgcat gacctcatgt gccagtgctg gcggaaggac cctgaggagc ggcccacttt
     2101 tgagtacctg caggccttcc tggaggacta cttcacctcg acagagcccc agtaccagcc
     2161 tggagagaac ctacgggatc caccggtcgc caccatggtg agcaagggcg aggagctgtt
     2221 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     2281 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     2341 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga gctggggcgt
     2401 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     2461 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     2521 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     2581 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaacg ccatccacgg
     2641 caacgtctat atcaccgccg acaagcagaa gaacggcatc aaggccaact tcggcctcaa
     2701 ctgcaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     2761 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaagctgag
     2821 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     2881 gatcactctc ggcatggacg agctgtacaa gtaaagcggc cgcgactcta gatcataatc
     2941 agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg
     3001 aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat
     3061 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     3121 tctagttgtg gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa
     3181 tattttgtta aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
     3241 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
     3301 tccagtttgg aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa
     3361 aaccgtctat cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg
     3421 gtcgaggtgc cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg
     3481 acggggaaag ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc
     3541 tagggcgctg gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa
     3601 tgcgccgcta cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     3661 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     3721 aatgcttcaa taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat
     3781 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3841 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     3901 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     3961 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4021 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4081 ggcttttttg gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt
     4141 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     4201 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     4261 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4321 actgcaagac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4381 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4441 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4501 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4561 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4621 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc
     4681 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4741 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4801 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4861 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4921 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4981 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     5041 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     5101 ttcgcccacc ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc
     5161 cgcgctatga cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat
     5221 aaacgcgggg ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg
     5281 ggccaatacg cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc
     5341 ccagggctcg cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat
     5401 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     5461 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     5521 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     5581 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     5641 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     5701 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     5761 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5821 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5881 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5941 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     6001 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     6061 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     6121 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     6181 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     6241 gccatgcatt agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat
     6301 ggagttccg
//
