LOCUS       sequence-420426-        7826 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of human Cdc45 C-terminally fused to mCerulean.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7826)
  TITLE     Pospiech Lab Plasmids 2023
REFERENCE   2  (bases 1 to 7826)
  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: pcDNA3-Cdc45-mCer
FEATURES             Location/Qualifiers
     source          1..7826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(50..69)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        241..620
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        621..824
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     775..795
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     869..888
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        869..887
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      2650..2659
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2656..2658,2659..2661,2662..3375)
                     /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(2701..2722)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2962..2981)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3309..3330
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(3385..3403)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3386..3403)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3423..3440)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3429..3653
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3699..4127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3786..3805)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3996..4017
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4136..4156)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4141..4470
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4321..4456
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4383..4402
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4537..5331
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4591..4610)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5201..5220
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5505..5626
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5542..5561)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5596..5615
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5675..5691)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5675..5691)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5688..5710)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5699..5715
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(5723..5729,5730..5747,5748..5753))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5768..5789
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5906..5923)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6077..6665)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6157..6176)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6836..7627,7628..7696))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7459..7478
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7697..7801)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt acaatctgct
       61 ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag gtcgctgagt
      121 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
      181 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      241 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
      301 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      361 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      421 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      481 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      541 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      601 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      661 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      721 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      781 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      841 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      901 ggtaccgagc tcggatccgc caccatgttc gtgtccgatt tccgcaaaga gttctacgag
      961 gtggtccaga gccagagggt ccttctcttc gtggcctcgg acgtggatgc tctgtgtgcg
     1021 tgcaagatcc ttcaggcctt gttccagtgt gaccacgtgc aatatacgct ggttccagtt
     1081 tctgggtggc aagaacttga aactgcattt cttgagcata aagaacagtt tcattatttt
     1141 attctcataa actgtggagc taatgtagac ctattggata ttcttcaacc tgatgaagac
     1201 actatattct ttgtgtgtga cacccatagg ccagtcaatg tcgtcaatgt atacaacgat
     1261 acccagatca aattactcat taaacaagat gatgaccttg aagttcccgc ctatgaagac
     1321 atcttcaggg atgaagagga ggatgaagag cattcaggaa atgacagtga tgggtcagag
     1381 ccttctgaga agcgcacacg gttagaagag gagatagtgg agcaaaccat gcggaggagg
     1441 cagcggcgag agtgggaggc ccggagaaga gacatcctct ttgactacga gcagtatgaa
     1501 tatcatggga catcgtcagc catggtgatg tttgagctgg cttggatgct gtccaaggac
     1561 ctgaatgaca tgctgtggtg ggccatcgtt ggactaacag accagtgggt gcaagacaag
     1621 atcactcaaa tgaaatacgt gactgatgtt ggtgtcctgc agcgccacgt ttcccgccac
     1681 aaccaccgga acgaggatga ggagaacaca ctctccgtgg actgcacacg gatctccttt
     1741 gagtatgacc tccgcctggt gctctaccag cactggtccc tccatgacag cctgtgcaac
     1801 accagctata ccgcagccag gttcaagctg tggtctgtgc atggacagaa gcggctccag
     1861 gagttccttg cagacatggg tcttcccctg aagcaggtga agcagaagtt ccaggccatg
     1921 gacatctcct tgaaggagaa tttgcgggaa atgattgaag agtctgcaaa taaatttggg
     1981 atgaaggaca tgcgcgtgca gactttcagc attcattttg ggttcaagca caagtttctg
     2041 gccagcgacg tggtctttgc caccatgtct ttgatggaga gccccgagaa ggatggctca
     2101 gggacagatc acttcatcca ggctctggac agcctctcca ggagtaacct ggacaagctg
     2161 taccatggcc tggaactcgc caagaagcag ctgcgagcca cccagcagac cattgccagc
     2221 tgcctttgca ccaacctcgt catctcccag gggcctttcc tgtactgctc tctcatggag
     2281 ggcactccag atgtcatgct gttctctagg ccggcatccc taagcctgct cagcaaacac
     2341 ctgctcaagt cctttgtgtg ttcgacaaag aaccggcgct gcaaactgct gcccctggtg
     2401 atggctgccc ccctgagcat ggagcatggc acagtgaccg tggtgggcat ccccccagag
     2461 accgacagct cggacaggaa gaactttttt gggagggcgt ttgagaaggc agcggaaagc
     2521 accagctccc ggatgctgca caaccatttt gacctctcag taattgagct gaaagctgag
     2581 gatcggagca agtttctgga cgcacttatt tccctcctgt ccctcgagca tgcatctaga
     2641 agatccatcg ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc
     2701 ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag
     2761 ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc
     2821 gtgccctggc ccaccctcgt gaccaccctg acctggggcg tgcagtgctt cgcccgctac
     2881 cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag
     2941 gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc
     3001 gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc
     3061 aacatcctgg ggcacaagct ggagtacaac gccatcagcg acaacgtcta tatcaccgcc
     3121 gacaagcaga agaacggcat caaggccaac ttcaagatcc gccacaacat cgaggacggc
     3181 agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg
     3241 ctgcccgaca accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag
     3301 cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac
     3361 gagctgtaca agtaagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc
     3421 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     3481 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     3541 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     3601 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     3661 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     3721 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     3781 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     3841 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     3901 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     3961 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     4021 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     4081 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat
     4141 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4201 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4261 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4321 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4381 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga
     4441 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc
     4501 ggatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4561 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4621 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4681 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     4741 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4801 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4861 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4921 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4981 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     5041 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     5101 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     5161 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     5221 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     5281 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     5341 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     5401 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     5461 tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag
     5521 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5581 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac
     5641 cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt
     5701 gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg
     5761 gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt
     5821 cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt
     5881 tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     5941 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     6001 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     6061 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     6121 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     6181 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     6241 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     6301 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     6361 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     6421 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     6481 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     6541 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     6601 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     6661 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     6721 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     6781 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     6841 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     6901 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     6961 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     7021 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     7081 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     7141 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     7201 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     7261 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     7321 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     7381 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     7441 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     7501 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     7561 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     7621 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     7681 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     7741 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     7801 gcacatttcc ccgaaaagtg ccacct
//
