LOCUS       sequence-420527-        6765 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  tet-inducible mammalian expression of human Cdc45 C-terminally fused
            to HA-Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6765)
  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 6765)
  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: pcDNA4/TO-Cdc45HA
FEATURES             Location/Qualifiers
     source          1..6765
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(46..65)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     771..791
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    822..840
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    843..861
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     871..895
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2708..2734
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(2775..2792)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2781..3005
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3051..3479
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3138..3157)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3348..3369
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3488..3508)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3493..3822
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3673..3808
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3735..3754
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3870..3917
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3936..4310
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4440..4561
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4477..4496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4531..4550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4610..4626)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4610..4626)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4623..4645)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4634..4650
                     /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(4658..4664,4665..4682,4683..4688))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4703..4724
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4841..4858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5012..5600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5092..5111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5771..6562,6563..6631))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSHIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6394..6413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6632..6736)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca gtgatagaga
      841 tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc agatcgcctg
      901 gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat ccagcctccg
      961 gactctagcg tttaaactta agcttggtac cgagctcgga tccgccacca tgttcgtgtc
     1021 cgatttccgc aaagagttct acgaggtggt ccagagccag agggtccttc tcttcgtggc
     1081 ctcggacgtg gatgctctgt gtgcgtgcaa gatccttcag gccttgttcc agtgtgacca
     1141 cgtgcaatat acgctggttc cagtttctgg gtggcaagaa cttgaaactg catttcttga
     1201 gcataaagaa cagtttcatt attttattct cataaactgt ggagctaatg tagacctatt
     1261 ggatattctt caacctgatg aagacactat attctttgtg tgtgacaccc ataggccagt
     1321 caatgtcgtc aatgtataca acgataccca gatcaaatta ctcattaaac aagatgatga
     1381 ccttgaagtt cccgcctatg aagacatctt cagggatgaa gaggaggatg aagagcattc
     1441 aggaaatgac agtgatgggt cagagccttc tgagaagcgc acacggttag aagaggagat
     1501 agtggagcaa accatgcgga ggaggcagcg gcgagagtgg gaggcccgga gaagagacat
     1561 cctctttgac tacgagcagt atgaatatca tgggacatcg tcagccatgg tgatgtttga
     1621 gctggcttgg atgctgtcca aggacctgaa tgacatgctg tggtgggcca tcgttggact
     1681 aacagaccag tgggtgcaag acaagatcac tcaaatgaaa tacgtgactg atgttggtgt
     1741 cctgcagcgc cacgtttccc gccacaacca ccggaacgag gatgaggaga acacactctc
     1801 cgtggactgc acacggatct cctttgagta tgacctccgc ctggtgctct accagcactg
     1861 gtccctccat gacagcctgt gcaacaccag ctataccgca gccaggttca agctgtggtc
     1921 tgtgcatgga cagaagcggc tccaggagtt ccttgcagac atgggtcttc ccctgaagca
     1981 ggtgaagcag aagttccagg ccatggacat ctccttgaag gagaatttgc gggaaatgat
     2041 tgaagagtct gcaaataaat ttgggatgaa ggacatgcgc gtgcagactt tcagcattca
     2101 ttttgggttc aagcacaagt ttctggccag cgacgtggtc tttgccacca tgtctttgat
     2161 ggagagcccc gagaaggatg gctcagggac agatcacttc atccaggctc tggacagcct
     2221 ctccaggagt aacctggaca agctgtacca tggcctggaa ctcgccaaga agcagctgcg
     2281 agccacccag cagaccattg ccagctgcct ttgcaccaac ctcgtcatct cccaggggcc
     2341 tttcctgtac tgctctctca tggagggcac tccagatgtc atgctgttct ctaggccggc
     2401 atccctaagc ctgctcagca aacacctgct caagtccttt gtgtgttcga caaagaaccg
     2461 gcgctgcaaa ctgctgcccc tggtgatggc tgcccccctg agcatggagc atggcacagt
     2521 gaccgtggtg ggcatccccc cagagaccga cagctcggac aggaagaact tttttgggag
     2581 ggcgtttgag aaggcagcgg aaagcaccag ctcccggatg ctgcacaacc attttgacct
     2641 ctcagtaatt gagctgaaag ctgaggatcg gagcaagttt ctggacgcac ttatttccct
     2701 cctgtcctac ccatacgacg tcccagacta cgcttagctc gagtctagag ggcccgttta
     2761 aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     2821 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     2881 ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca
     2941 ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc
     3001 tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg
     3061 tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc
     3121 cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg
     3181 ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg
     3241 gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg
     3301 atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt
     3361 ccaaactgga acaacactca accctatctc ggtctattct tttgatttat aagggatttt
     3421 gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta
     3481 attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga
     3541 agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc
     3601 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc
     3661 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     3721 tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag
     3781 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt
     3841 atatccattt tcggatctga tcagcacgtg ttgacaatta atcatcggca tagtatatcg
     3901 gcatagtata atacgacaag gtgaggaact aaaccatggc caagttgacc agtgccgttc
     3961 cggtgctcac cgcgcgcgac gtcgccggag cggtcgagtt ctggaccgac cggctcgggt
     4021 tctcccggga cttcgtggag gacgacttcg ccggtgtggt ccgggacgac gtgaccctgt
     4081 tcatcagcgc ggtccaggac caggtggtgc cggacaacac cctggcctgg gtgtgggtgc
     4141 gcggcctgga cgagctgtac gccgagtggt cggaggtcgt gtccacgaac ttccgggacg
     4201 cctccgggcc ggccatgacc gagatcggcg agcagccgtg ggggcgggag ttcgccctgc
     4261 gcgacccggc cggcaactgc gtgcacttcg tggccgagga gcaggactga cacgtgctac
     4321 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     4381 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4561 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     4621 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     4681 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     4741 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     4801 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     4861 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     4921 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     4981 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5041 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     5101 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     5161 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     5221 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     5281 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     5341 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     5401 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     5461 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     5521 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     5581 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     5641 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     5701 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     5761 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     5821 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatatgg gagggcttac
     5881 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     5941 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6001 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6061 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     6121 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     6181 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     6241 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     6301 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     6361 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     6421 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     6481 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     6541 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     6601 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     6661 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     6721 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacg
//
