LOCUS       sequence-233119-        5036 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 5036)
  TITLE     Davidson Cyan Fluorescent Proteins
REFERENCE   2  (bases 1 to 5036)
  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-H4-23
FEATURES             Location/Qualifiers
     source          1..5036
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        86..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    941..993
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      1001..1010
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1007..1009,1010..1012,1013..1726)
                     /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(1052..1073)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1313..1332)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1660..1681
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1849..1970
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1886..1905)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1940..1959
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1977..2432)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2114..2135)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2326..2345
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2459..2563
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2565..2922
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2565..2756
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2588..2608)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2773..2908
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2957..3751
                     /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             2957..3751
                     /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(3011..3030)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3621..3640
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3939..3958)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3983..4030
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4359..4947
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4848..4867
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggataaccgt attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      601 tgaaccgtca gatccgctag ccgccaccat gtctggccgc ggcaaaggcg ggaagggtct
      661 tggcaaaggc ggcgctaagc gccaccgtaa agtactgcgc gacaatatcc agggcatcac
      721 caagccggcc atccggcgcc ttgctcgccg cggcggcgtg aagcgcatct ccggcctcat
      781 ctacgaggag actcgcgggg tgctgaaggt gttcctggag aacgtgatcc gggacgccgt
      841 gacctataca gagcacgcca agcgcaagac ggtcaccgcc atggatgtgg tctacgcgct
      901 caagcgccag ggccgcaccc tctacggttt cggtggggat ctcgagctca agcttcgaat
      961 tctgcagtcg acggtaccgc gggcccggga tccaccggtc gccaccatgg tgagcaaggg
     1021 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1081 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1141 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1201 gacctggggc gtgcagtgct tcgcccgcta ccccgaccac atgaagcagc acgacttctt
     1261 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1321 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1381 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1441 cgccatcagc gacaacgtct atatcaccgc cgacaagcag aagaacggca tcaaggccaa
     1501 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1561 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1621 gtccaagctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1681 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaaagcg gccgcgactc
     1741 tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa aaacctccca
     1801 cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa cttgtttatt
     1861 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     1921 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta aggcgtaaat
     1981 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     2041 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     2101 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     2161 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     2221 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     2281 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     2341 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     2401 cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg
     2461 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2521 ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg cggaaagaac
     2581 cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     2641 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     2701 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     2761 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     2821 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     2881 aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc aagagacagg
     2941 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     3001 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     3061 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     3121 tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     3181 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     3241 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     3301 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     3361 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     3421 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     3481 ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     3541 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     3601 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     3661 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3721 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3781 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3841 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3901 atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata
     3961 ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg
     4021 tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg
     4081 agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt
     4141 tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca
     4201 ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     4261 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     4321 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     4381 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     4441 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     4501 actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     4561 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     4621 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     4681 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     4741 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     4801 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4861 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4921 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4981 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgt
//
