LOCUS       sequence-271842-        5719 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of CENP-C.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5719)
  AUTHORS   Swartz SZ, McKay LS, Su KC, Bury L, Padeganeh A, Maddox PS, Knouse 
            KA, Cheeseman IM
  TITLE     Quiescent Cells Actively Replenish CENP-A Nucleosomes to Maintain 
            Centromere Identity and Proliferative Potential.
  JOURNAL   Dev Cell. 2019 Oct 7;51(1):35-48.e7. doi: 
            10.1016/j.devcel.2019.07.016. Epub 2019 Aug 15.
  PUBMED    31422918
REFERENCE   2  (bases 1 to 5719)
  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"; doi: 
            "10.1016/j.devcel.2019.07.016"; journalName: "Dev Cell"; date: 
            "2019-10-7- 7"; volume: "51"; issue: "1"; pages: "35-48.e7"
COMMENT     Sequence Label: pZS077
FEATURES             Location/Qualifiers
     source          1..5719
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..173
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        246..323
                     /gene="lacI"
                     /label=lacI promoter
     CDS             324..1406
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(343..362)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1455..1460,1461..1478,1479..1485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1493..1509
                     /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)."
     primer_bind     1517..1533
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1517..1533
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1529..2056
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1549..1566)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1549..1565)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1558..1580)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2158..2163,2164..2179,2180..2186)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2194..2210
                     /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)."
     CDS             2233..2886
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2844..2866
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2893..2916
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3744..3766)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3911..3929)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3997..4101
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4102..4170,4171..4962)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(4320..4339)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(5133..5719,1..2)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5622..5641
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
       61 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      121 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      181 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
      241 attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
      301 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
      361 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
      421 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
      481 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
      541 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
      601 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
      661 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
      721 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
      781 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
      841 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
      901 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
      961 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
     1021 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
     1081 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     1141 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     1201 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     1261 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca
     1321 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     1381 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     1441 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     1501 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
     1561 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1621 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1681 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     1741 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     1801 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
     1861 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     1921 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt
     1981 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     2041 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     2101 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     2161 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     2221 gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc
     2281 actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat
     2341 gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct
     2401 tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct
     2461 gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa
     2521 ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa
     2581 actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt
     2641 ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat
     2701 gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta
     2761 gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc
     2821 aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct
     2881 ccaaaatcgg atctggaagt tctgttccag gggcccctgg gatccatgtc agcagaaaag
     2941 gaaagaagat accaggaact ctacaagaat cgctttgtga atcccattga tcaaggcatt
     3001 ggcaggcgta caggatgggt gcttaacaag gaggtgcgta aggacagtga aggctttgag
     3061 ttatttgatg actactggtc agattctggt gaggagagtg ttgatgactt gtcccttcat
     3121 gccaacaaag aaaataaagg gagaccgtac aagctgagtt ctcccataaa gcacatgcca
     3181 gactttactc cgagttctcg ggccagcaca gatgtggaac ccttgtctga tgttgtctcc
     3241 aaaacagtca cttcaaattc cactctgacc aaagcttcaa ataatctttc aaagattgcc
     3301 aagtcagtta aagcggatca ggctaaaact aacgaagagc cgttggctat agagagggag
     3361 aatggcataa ccagggagga aggtgatgct ggagatgaag cgacaacacc aggccgagag
     3421 gaaggcggta ttcaagagtc agacatcact ggaagcatta ttaaaggagg aaattctttc
     3481 caggttccca actctgcact tcaggaattc aagaccaaga caaggttaag ctttggtaat
     3541 gacgttgacg atacaccaaa aaggagatcc acaaatggaa catcatcaag agcagtcggg
     3601 agtcaaagcg atgaggagag tgtctcatcg gataccacct tgaaaggatc tcagtctgat
     3661 gtgaacttgg acggctaact cgagcggccg catcgtgact gactgacgat ctgcctcgcg
     3721 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct
     3781 tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc
     3841 gggtgtcggg gcgcagccat gacccagtca cgtagcgata gcggagtgta taattcttga
     3901 agacgaaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat aataatggtt
     3961 tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     4021 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4081 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4141 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4201 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4261 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4321 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     4381 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4441 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4501 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4561 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     4621 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     4681 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     4741 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     4801 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     4861 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     4921 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     4981 tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag
     5041 atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     5101 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     5161 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     5221 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     5281 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     5341 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     5401 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     5461 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     5521 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     5581 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5641 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5701 ggggggcgga gcctatgga
//
