LOCUS       sequence-170848-        5139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Nucleus/Histones, Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5139)
  TITLE     Davidson Cyan Fluorescent Proteins
REFERENCE   2  (bases 1 to 5139)
  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"
COMMENT     Sequence Label: mCerulean-H3.3-N-14
FEATURES             Location/Qualifiers
     source          1..5139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      265..274
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      715..724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(721..723,724..726,727..1440)
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Rizzo et al., 
                     2004)"
                     /label=mCerulean
                     /note=" mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(766..787)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1027..1046)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1374..1395
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1563..1684
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1600..1619)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1654..1673
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1691..2146)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1828..1849)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2040..2059
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2173..2277
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2279..2636
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2279..2470
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2302..2322)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2487..2622
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2671..3465
                     /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             2671..3465
                     /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(2725..2744)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3335..3354
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3653..3672)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3697..3744
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4073..4661
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4562..4581
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        4836..5139
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct cgagctcaag cttcgccacc atggcccgaa ccaagcagac cgctaggaag
      301 tccaccggtg ggaaagcccc ccgcaaacag ctggccacca aggcggctcg gaaaagcgcg
      361 ccctctaccg gcggggtgaa gaagcctcac cgctacaggc cagggaccgt ggctctgaga
      421 gagatccgtc gttaccagaa atcgactgag ctgctcatcc ggaagctgcc attccagaga
      481 ttggtgaggg agatcgccca ggatttcaaa accgacttga ggtttcaaag tgcagccatc
      541 ggtgcccttc aggaggctag cgaagcatac ctggtggggt tgtttgaaga taccaatctg
      601 tgtgccatcc acgccaagag agtcaccatc atgcccaaag acatccagtt ggctcgccgg
      661 atacgggggg agagagctgg gtccggaagc gcaggcagtg gggatccacc ggtcgccacc
      721 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      781 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      841 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      901 ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc gctaccccga ccacatgaag
      961 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     1021 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     1081 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     1141 aagctggagt acaacgccat cagcgacaac gtctatatca ccgccgacaa gcagaagaac
     1201 ggcatcaagg ccaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     1261 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     1321 tacctgagca cccagtccaa gctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     1381 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     1441 agcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt
     1501 taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg
     1561 ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     1621 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     1681 cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     1741 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     1801 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     1861 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     1921 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     1981 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     2041 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     2101 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt
     2161 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2221 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct
     2281 gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct
     2341 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa
     2401 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     2461 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     2521 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     2581 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc
     2641 gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt
     2701 tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc
     2761 tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag
     2821 accgacctgt ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg
     2881 gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac
     2941 tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc
     3001 gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc
     3061 tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc
     3121 ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg
     3181 ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat
     3241 gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc
     3301 cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa
     3361 gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat
     3421 tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt
     3481 tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3541 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3601 agcgcgggga tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac
     3661 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     3721 cgcacggtgt tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg
     3781 tcgatacccc accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac
     3841 cccacccccc aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc
     3901 ctgccatagc ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat
     3961 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     4021 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4081 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4141 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4201 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     4261 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4321 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4381 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4441 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4501 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4561 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4621 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4681 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4741 ctgattctgt ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg
     4801 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
     4861 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
     4921 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
     4981 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
     5041 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
     5101 tggcagtaca tctacgtatt agtcatcgct attaccatg
//
