LOCUS       sequence-420495-        6430 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human Cdc45 C-terminally fused to mCherry.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6430)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 6430)
  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: pCdc45-mCherry
FEATURES             Location/Qualifiers
     source          1..6430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    625..681
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2395..2404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2401..3111
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2546..2564)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2814..2833
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3234..3355
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3271..3290)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3325..3344
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3362..3817)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3499..3520)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3711..3730
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3844..3948
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3950..4307
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3950..4141
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3973..3993)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4158..4293
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4342..5136
                     /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             4342..5136
                     /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(4396..4415)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5006..5025
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5324..5343)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5368..5415
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5744..6332
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6233..6252
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tattaccgcc atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctggttta gtgaaccgtc
      601 agatccgcta gcgctaccgg actcagatct cgagctcaag cttcgaattc tgcagtcgac
      661 ggtaccgcgg gcccgggatc cgccaccatg ttcgtgtccg atttccgcaa agagttctac
      721 gaggtggtcc agagccagag ggtccttctc ttcgtggcct cggacgtgga tgctctgtgt
      781 gcgtgcaaga tccttcaggc cttgttccag tgtgaccacg tgcaatatac gctggttcca
      841 gtttctgggt ggcaagaact tgaaactgca tttcttgagc ataaagaaca gtttcattat
      901 tttattctca taaactgtgg agctaatgta gacctattgg atattcttca acctgatgaa
      961 gacactatat tctttgtgtg tgacacccat aggccagtca atgtcgtcaa tgtatacaac
     1021 gatacccaga tcaaattact cattaaacaa gatgatgacc ttgaagttcc cgcctatgaa
     1081 gacatcttca gggatgaaga ggaggatgaa gagcattcag gaaatgacag tgatgggtca
     1141 gagccttctg agaagcgcac acggttagaa gaggagatag tggagcaaac catgcggagg
     1201 aggcagcggc gagagtggga ggcccggaga agagacatcc tctttgacta cgagcagtat
     1261 gaatatcatg ggacatcgtc agccatggtg atgtttgagc tggcttggat gctgtccaag
     1321 gacctgaatg acatgctgtg gtgggccatc gttggactaa cagaccagtg ggtgcaagac
     1381 aagatcactc aaatgaaata cgtgactgat gttggtgtcc tgcagcgcca cgtttcccgc
     1441 cacaaccacc ggaacgagga tgaggagaac acactctccg tggactgcac acggatctcc
     1501 tttgagtatg acctccgcct ggtgctctac cagcactggt ccctccatga cagcctgtgc
     1561 aacaccagct ataccgcagc caggttcaag ctgtggtctg tgcatggaca gaagcggctc
     1621 caggagttcc ttgcagacat gggtcttccc ctgaagcagg tgaagcagaa gttccaggcc
     1681 atggacatct ccttgaagga gaatttgcgg gaaatgattg aagagtctgc aaataaattt
     1741 gggatgaagg acatgcgcgt gcagactttc agcattcatt ttgggttcaa gcacaagttt
     1801 ctggccagcg acgtggtctt tgccaccatg tctttgatgg agagccccga gaaggatggc
     1861 tcagggacag atcacttcat ccaggctctg gacagcctct ccaggagtaa cctggacaag
     1921 ctgtaccatg gcctggaact cgccaagaag cagctgcgag ccacccagca gaccattgcc
     1981 agctgccttt gcaccaacct cgtcatctcc caggggcctt tcctgtactg ctctctcatg
     2041 gagggcactc cagatgtcat gctgttctct aggccggcat ccctaagcct gctcagcaaa
     2101 cacctgctca agtcctttgt gtgttcgaca aagaaccggc gctgcaaact gctgcccctg
     2161 gtgatggctg cccccctgag catggagcat ggcacagtga ccgtggtggg catcccccca
     2221 gagaccgaca gctcggacag gaagaacttt tttgggaggg cgtttgagaa ggcagcggaa
     2281 agcaccagct cccggatgct gcacaaccat tttgacctct cagtaattga gctgaaagct
     2341 gaggatcgga gcaagtttct ggacgcactt atttccctcc tgtccgaacc ggtcgccacc
     2401 atggtgagca agggcgagga ggataacatg gccatcatca aggagttcat gcgcttcaag
     2461 gtgcacatgg agggctccgt gaacggccac gagttcgaga tcgagggcga gggcgagggc
     2521 cgcccctacg agggcaccca gaccgccaag ctgaaggtga ccaagggtgg ccccctgccc
     2581 ttcgcctggg acatcctgtc ccctcagttc atgtacggct ccaaggccta cgtgaagcac
     2641 cccgccgaca tccccgacta cttgaagctg tccttccccg agggcttcaa gtgggagcgc
     2701 gtgatgaact tcgaggacgg cggcgtggtg accgtgaccc aggactcctc cctgcaggac
     2761 ggcgagttca tctacaaggt gaagctgcgc ggcaccaact tcccctccga cggccccgta
     2821 atgcagaaga agaccatggg ctgggaggcc tcctccgagc ggatgtaccc cgaggacggc
     2881 gccctgaagg gcgagatcaa gcagaggctg aagctgaagg acggcggcca ctacgacgct
     2941 gaggtcaaga ccacctacaa ggccaagaag cccgtgcagc tgcccggcgc ctacaacgtc
     3001 aacatcaagt tggacatcac ctcccacaac gaggactaca ccatcgtgga acagtacgaa
     3061 cgcgccgagg gccgccactc caccggcggc atggacgagc tgtacaagta aagcggccgc
     3121 gactctagat cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc
     3181 tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt
     3241 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3301 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaaggcg
     3361 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     3421 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     3481 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     3541 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     3601 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     3661 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     3721 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     3781 acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt tcggggaaat
     3841 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     3901 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa
     3961 agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag
     4021 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     4081 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     4141 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     4201 atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat
     4261 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat cgatcaagag
     4321 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     4381 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     4441 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4501 tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct ggccacgacg
     4561 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4621 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4681 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4741 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4801 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4861 ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4921 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4981 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     5041 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     5101 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     5161 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     5221 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     5281 atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca cggaaggaga
     5341 caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa acgcacggtg
     5401 ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct gtcgataccc
     5461 caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca ccccaccccc
     5521 caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag
     5581 cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     5641 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     5701 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     5761 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     5821 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     5881 caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     5941 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     6001 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     6061 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     6121 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     6181 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     6241 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     6301 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     6361 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     6421 tggataaccg
//
