LOCUS       sequence-420341-        6392 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human Cdc45 C-terminally fused to EGFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6392)
  AUTHORS   Kohler C, Koalick D, Fabricius A, Parplys AC, Borgmann K, Pospiech 
            H, Grosse F
  TITLE     Cdc45 is limiting for replication initiation in humans.
  JOURNAL   Cell Cycle. 2016;15(7):974-85. doi: 10.1080/15384101.2016.1152424.
  PUBMED    26919204
REFERENCE   2  (bases 1 to 6392)
  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: "Cell 
            Cycle."; date: "2016"; volume: "15(7):974-85. doi"; pages: " 
            10.1080/15384101.2016.1152424"
COMMENT     Sequence Label: pEGFP-N1-Cdc45
FEATURES             Location/Qualifiers
     source          1..6392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        36..339
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        340..543
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     494..514
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2307..2316
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2313..2315,2316..2318,2319..3032)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2313..2315,2316..2318,2319..3029)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2358..2379)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2619..2638)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2966..2987
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3155..3276
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3192..3211)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3246..3265
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3283..3738)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3420..3441)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3632..3651
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3765..3869
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3871..4228
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3871..4062
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3894..3914)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4079..4214
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4263..5057
                     /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             4263..5057
                     /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(4317..4336)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4927..4946
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5245..5264)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5289..5336
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5665..6253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6154..6173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc
       61 ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc
      121 atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact
      181 gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat
      241 gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact
      301 tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac
      361 atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac
      421 gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac
      481 tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga
      541 gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc gagatgttcg
      601 tgtccgattt ccgcaaagag ttctacgagg tggtccagag ccagagggtc cttctcttcg
      661 tggcctcgga cgtggatgct ctgtgtgcgt gcaagatcct tcaggccttg ttccagtgtg
      721 accacgtgca atatacgctg gttccagttt ctgggtggca agaacttgaa actgcatttc
      781 ttgagcataa agaacagttt cattatttta ttctcataaa ctgtggagct aatgtagacc
      841 tattggatat tcttcaacct gatgaagaca ctatattctt tgtgtgtgac acccataggc
      901 cagtcaatgt cgtcaatgta tacaacgata cccagatcaa attactcatt aaacaagatg
      961 atgaccttga agttcccgcc tatgaagaca tcttcaggga tgaagaggag gatgaagagc
     1021 attcaggaaa tgacagtgat gggtcagagc cttctgagaa gcgcacacgg ttagaagagg
     1081 agatagtgga gcaaaccatg cggaggaggc agcggcgaga gtgggaggcc cggagaagag
     1141 acatcctctt tgactacgag cagtatgaat atcatgggac atcgtcagcc atggtgatgt
     1201 ttgagctggc ttggatgctg tccaaggacc tgaatgacat gctgtggtgg gccatcgttg
     1261 gactaacaga ccagtgggtg caagacaaga tcactcaaat gaaatacgtg actgatgttg
     1321 gtgtcctgca gcgccacgtt tcccgccaca accaccggaa cgaggatgag gagaacacac
     1381 tctccgtgga ctgcacacgg atctcctttg agtatgacct ccgcctggtg ctctaccagc
     1441 actggtccct ccatgacagc ctgtgcaaca ccagctatac cgcagccagg ttcaagctgt
     1501 ggtctgtgca tggacagaag cggctccagg agttccttgc agacatgggt cttcccctga
     1561 agcaggtgaa gcagaagttc caggccatgg acatctcctt gaaggagaat ttgcgggaaa
     1621 tgattgaaga gtctgcaaat aaatttggga tgaaggacat gcgcgtgcag actttcagca
     1681 ttcattttgg gttcaagcac aagtttctgg ccagcgacgt ggtctttgcc accatgtctt
     1741 tgatggagag ccccgagaag gatggctcag ggacagatca cttcatccag gctctggaca
     1801 gcctctccag gagtaacctg gacaagctgt accatggcct ggaactcgcc aagaagcagc
     1861 tgcgagccac ccagcagacc attgccagct gcctttgcac caacctcgtc atctcccagg
     1921 ggcctttcct gtactgctct ctcatggagg gcactccaga tgtcatgctg ttctctaggc
     1981 cggcatccct aagcctgctc agcaaacacc tgctcaagtc ctttgtgtgt tcgacaaaga
     2041 accggcgctg caaactgctg cccctggtga tggctgcccc cctgagcatg gagcatggca
     2101 cagtgaccgt ggtgggcatc cccccagaga ccgacagctc ggacaggaag aacttttttg
     2161 ggagggcgtt tgagaaggca gcggaaagca ccagctcccg gatgctgcac aaccattttg
     2221 acctctcagt aattgagctg aaagctgagg atcggagcaa gtttctggac gcacttattt
     2281 ccctcctgtc ccgggatcca ccggtcgcca ccatggtgag caagggcgag gagctgttca
     2341 ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg
     2401 tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca
     2461 ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc
     2521 agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc
     2581 ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc
     2641 gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg
     2701 acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca
     2761 acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc
     2821 acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg
     2881 gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca
     2941 aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga
     3001 tcactctcgg catggacgag ctgtacaagt aaagcggccg cgactctaga tcataatcag
     3061 ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa
     3121 cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg
     3181 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     3241 tagttgtggt ttgtccaaac tcatcaatgt atcttaaggc gtaaattgta agcgttaata
     3301 ttttgttaaa attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg
     3361 aaatcggcaa aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc
     3421 cagtttggaa caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa
     3481 ccgtctatca gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt
     3541 cgaggtgccg taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac
     3601 ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta
     3661 gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg
     3721 cgccgctaca gggcgcgtca ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     3781 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     3841 tgcttcaata atattgaaaa aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt
     3901 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3961 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     4021 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     4081 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     4141 tatttatgca gaggccgagg ccgcctcggc ctctgagcta ttccagaagt agtgaggagg
     4201 cttttttgga ggcctaggct tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc
     4261 gcatgattga acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat
     4321 tcggctatga ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt
     4381 cagcgcaggg gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac
     4441 tgcaagacga ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg
     4501 tgctcgacgt tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc
     4561 aggatctcct gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa
     4621 tgcggcggct gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc
     4681 gcatcgagcg agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg
     4741 aagagcatca ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg agcatgcccg
     4801 acggcgagga tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa
     4861 atggccgctt ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg
     4921 acatagcgtt ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct
     4981 tcctcgtgct ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc
     5041 ttgacgagtt cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa
     5101 cctgccatca cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5161 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5221 cgcccaccct agggggaggc taactgaaac acggaaggag acaataccgg aaggaacccg
     5281 cgctatgacg gcaataaaaa gacagaataa aacgcacggt gttgggtcgt ttgttcataa
     5341 acgcggggtt cggtcccagg gctggcactc tgtcgatacc ccaccgagac cccattgggg
     5401 ccaatacgcc cgcgtttctt ccttttcccc accccacccc ccaagttcgg gtgaaggccc
     5461 agggctcgca gccaacgtcg gggcggcagg ccctgccata gcctcaggtt actcatatat
     5521 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5581 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6301 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6361 catgcattag ttattaatag taatcaatta cg
//
