LOCUS       sequence-338677-        4850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Endoplasmic Reticulum, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4850)
  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 4850)
  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-CytERM-N-17
FEATURES             Location/Qualifiers
     source          1..4850
                     /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"
     regulatory      730..739
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(736..738,739..741,742..1455)
                     /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(781..802)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1042..1061)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1389..1410
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1578..1699
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1615..1634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1669..1688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1706..2161)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1843..1864)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2055..2074
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2188..2292
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2294..2651
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2294..2485
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2317..2337)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2502..2637
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2686..3480
                     /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             2686..3480
                     /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(2740..2759)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3350..3369
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3668..3687)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3712..3759
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4088..4676
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4577..4596
                     /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 atctggtacc ccttgcagct tccagttgtc aggaagagaa ggctcccatg
      601 gatcctgtgg tggtgctggg gctctgtctc tcctgtttgc ttctcctttc actctggaaa
      661 cagagctatg ggggagggaa gcttcgaatt ctgcagtcga cggtaccgcg ggcccgggat
      721 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
      781 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
      841 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
      901 gtgccctggc ccaccctcgt gaccaccctg agctggggcg tgcagtgctt cgcccgctac
      961 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     1021 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     1081 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     1141 aacatcctgg ggcacaagct ggagtacaac gccatccacg gcaacgtcta tatcaccgcc
     1201 gacaagcaga agaacggcat caaggccaac ttcggcctca actgcaacat cgaggacggc
     1261 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     1321 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     1381 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     1441 gagctgtaca agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag
     1501 aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga
     1561 atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     1621 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     1681 aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg
     1741 ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct
     1801 tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt
     1861 ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat
     1921 ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca
     1981 ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac
     2041 gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta
     2101 gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg
     2161 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     2221 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     2281 aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg
     2341 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2401 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2461 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2521 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2581 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2641 gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2701 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2761 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2821 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg
     2881 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2941 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     3001 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     3061 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     3121 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     3181 gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc
     3241 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     3301 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3361 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3421 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3481 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3541 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3601 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga
     3661 ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa
     3721 aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc
     3781 agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt
     3841 cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg
     3901 tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag attgatttaa
     3961 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     4021 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     4081 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     4141 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     4201 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     4261 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     4321 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     4381 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     4441 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     4501 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     4561 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     4621 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     4681 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct
     4741 ttcctgcgtt atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa
     4801 tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
//
