LOCUS       sequence-478928-        5995 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Note- This plasmid contains Keratin 18. Localization: Cytokeratin, 
            Excitation: 433, Emission: 475.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5995)
  AUTHORS   Rizzo MA, Davidson MW, Piston DW
  TITLE     Fluorescent protein tracking and detection: fluorescent protein 
            structure and color variants.
  JOURNAL   Cold Spring Harb Protoc. 2009 Dec;2009(12):pdb.top63. doi: 
            10.1101/pdb.top63.
  PUBMED    20150100
REFERENCE   2  (bases 1 to 5995)
  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: "Cold Spring
            Harb Protoc."; date: "2009-12"; volume: "2009(12):pdb.top63. doi"; 
            pages: " 10.1101/pdb.top63"
COMMENT     Sequence Label: mCerulean-Keratin-17
FEATURES             Location/Qualifiers
     source          1..5995
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..40
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1436..1445
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1442..1444,1445..1447,1448..2161)
                     /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(1487..1508)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1748..1767)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2095..2116
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2284..2405
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2321..2340)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2375..2394
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2412..2867)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2549..2570)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2761..2780
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2894..2998
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3000..3357
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3000..3191
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3023..3043)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3208..3343
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3392..4186
                     /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             3392..4186
                     /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(3446..3465)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4056..4075
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4374..4393)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4418..4465
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4794..5382
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5283..5302
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        5557..5860
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        join(5861..5995,1..69)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
ORIGIN
        1 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
       61 agcagagctg gtttagtgaa ccgtcagatc cgctagcacc atgagcttca ccactcgctc
      121 caccttctcc accaactacc ggtccctggg ctctgtccag gcgcccagct acggcgcccg
      181 gccggtcagc agcgcggcca gcgtctatgc aggcgctggg ggctctggtt cccggatctc
      241 cgtgtcccgc tccaccagct tcaggggcgg catggggtcc gggggcctgg ccaccgggat
      301 agccgggggt ctggcaggaa tgggaggcat ccagaacgag aaggagacca tgcaaagcct
      361 gaacgaccgc ctggcctctt acctggacag agtgaggagc ctggagaccg agaaccggag
      421 gctggagagc aaaatccggg agcacttgga gaagaaggga ccccaggtca gagactggag
      481 ccattacttc aagatcatcg aggacctgag ggctcagatc ttcgcaaata ctgtggacaa
      541 tgcccgcatc gttctgcaga ttgacaatgc ccgtcttgct gctgatgact ttagagtcaa
      601 gtatgagaca gagctggcca tgcgccagtc tgtggagaac gacatccatg ggctccgcaa
      661 ggtcattgat gacaccaata tcacacgact gcagctggag acagagatcg aggctctcaa
      721 ggaggagctg ctcttcatga agaagaacca cgaagaggaa gtaaaaggcc tacaagccca
      781 gattgccagc tctgggttga ccgtggaggt agatgccccc aaatctcagg acctcgccaa
      841 gatcatggca gacatccggg cccaatatga cgagctggct cggaagaacc gagaggagct
      901 agacaagtac tggtctcagc agattgagga gagcaccaca gtgctcacca cacagtctgc
      961 tgaggttgga gctgctgaga cgacgctcac agagctgaga cgtacagtcc agtccttgga
     1021 gatcgacctg gactccatga gaaatctgaa ggccagcttg gagaacagcc tgagggaggt
     1081 ggaggcccgc tacgccctac agatggagca gctcaacggg atcctgctgc accttgagtc
     1141 agagctggca cagacccggg cagagggaca gcgccaggcc caggagtatg aggccctgct
     1201 gaacatcaag gtcaagctgg aggctgagat cgccacctac cgccgcctgc tggaagatgg
     1261 cgaggacttt aatcttggtg atgccttgga cagcagcaac tccatgcaaa ccatccaaaa
     1321 gaccaccacc cgccggatag tggatggcaa agtggtgtct gagaccaatg acaccaaagt
     1381 tctgaggcat cgaattctgc agtcgacggt accgcgggcc cgggatccac cggtcgccac
     1441 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     1501 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     1561 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     1621 cctcgtgacc accctgacct ggggcgtgca gtgcttcgcc cgctaccccg accacatgaa
     1681 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     1741 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     1801 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     1861 caagctggag tacaacgcca tcagcgacaa cgtctatatc accgccgaca agcagaagaa
     1921 cggcatcaag gccaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     1981 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     2041 ctacctgagc acccagtcca agctgagcaa agaccccaac gagaagcgcg atcacatggt
     2101 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     2161 aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct
     2221 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2281 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2341 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2401 tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     2461 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     2521 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     2581 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2641 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2701 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2761 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2821 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     2881 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     2941 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3001 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3061 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3121 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3181 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3241 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3301 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3361 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3421 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     3481 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     3541 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     3601 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     3661 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     3721 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     3781 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     3841 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     3901 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     3961 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4021 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4081 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4141 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4201 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4261 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4321 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4381 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4441 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4501 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4561 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4621 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     4681 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     4741 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     4801 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     4861 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     4921 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     4981 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5041 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5101 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5161 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5221 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5281 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5341 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5401 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     5461 cctgattctg tggataaccg tattaccgcc atgcattagt tattaatagt aatcaattac
     5521 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg
     5581 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc
     5641 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac
     5701 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa
     5761 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac
     5821 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta
     5881 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga
     5941 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac ttttcaaaat gtcgt
//
