LOCUS       sequence-271845-        5188 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For in vitro transcription of Miniata CENP-A.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5188)
  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 5188)
  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: pZS039
FEATURES             Location/Qualifiers
     source          1..5188
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..53
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        126..203
                     /gene="lacI"
                     /label=lacI promoter
     CDS             204..1286
                     /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(223..242)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1335..1340,1341..1358,1359..1365)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1373..1389
                     /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     1397..1413
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1397..1413
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1409..1936
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1429..1446)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1429..1445)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1438..1460)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(2038..2043,2044..2059,2060..2066)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2074..2090
                     /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             2113..2766
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2724..2746
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2773..2796
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     primer_bind     complement(3093..3115)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3260..3278)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3346..3450
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3451..3519,3520..4311)
                     /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(3669..3688)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4482..5070
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4971..4990
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
       61 aagagcgcct gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa
      121 attccgacac catcgaatgg cgcaaaacct ttcgcggtat ggcatgatag cgcccggaag
      181 agagtcaatt cagggtggtg aatgtgaaac cagtaacgtt atacgatgtc gcagagtatg
      241 ccggtgtctc ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga
      301 aaacgcggga aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg
      361 cacaacaact ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc
      421 tgcacgcgcc gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca
      481 gcgtggtggt gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca
      541 atcttctcgc gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg
      601 ccattgctgt ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc
      661 agacacccat caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc
      721 atctggtcgc attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct
      781 cggcgcgtct gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga
      841 tagcggaacg ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc
      901 tgaatgaggg catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg
      961 caatgcgcgc cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat
     1021 acgacgatac cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt
     1081 ttcgcctgct ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg
     1141 tgaagggcaa tcagctgttg cccgtctcac tggtgaaaag aaaaaccacg ctagcgccca
     1201 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     1261 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtaagtta gctcactcat
     1321 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     1381 ggataacaat ttcacacagg aaacagctat gaccatgatt acggattcac tggccgtcgt
     1441 tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca
     1501 tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca
     1561 gttgcgcagc ctgaatggcg aatggcgctt tgcctggttt ccggcaccag aagcggtgcc
     1621 ggaaagctgg ctggagtgcg atcttcctga ggccgatact gtcgtcgtcc cctcaaactg
     1681 gcagatgcac ggttacgatg cgcccatcta caccaacgtg acctatccca ttacggtcaa
     1741 tccgccgttt gttcccacgg agaatccgac gggttgttac tcgctcacat ttaatgttga
     1801 tgaaagctgg ctacaggaag gccagacgcg aattattttt gatggcgttg gaattagctt
     1861 atcgactgca cggtgcacca atgcttctgg cgtcaggcag ccatcggaag ctgtggtatg
     1921 gctgtgcagg tcgtaaatca ctgcataatt cgtgtcgctc aaggcgcact cccgttctgg
     1981 ataatgtttt ttgcgccgac atcataacgg ttctggcaaa tattctgaaa tgagctgttg
     2041 acaattaatc atcggctcgt ataatgtgtg gaattgtgag cggataacaa tttcacacag
     2101 gaaacagtat tcatgtcccc tatactaggt tattggaaaa ttaagggcct tgtgcaaccc
     2161 actcgacttc ttttggaata tcttgaagaa aaatatgaag agcatttgta tgagcgcgat
     2221 gaaggtgata aatggcgaaa caaaaagttt gaattgggtt tggagtttcc caatcttcct
     2281 tattatattg atggtgatgt taaattaaca cagtctatgg ccatcatacg ttatatagct
     2341 gacaagcaca acatgttggg tggttgtcca aaagagcgtg cagagatttc aatgcttgaa
     2401 ggagcggttt tggatattag atacggtgtt tcgagaattg catatagtaa agactttgaa
     2461 actctcaaag ttgattttct tagcaagcta cctgaaatgc tgaaaatgtt cgaagatcgt
     2521 ttatgtcata aaacatattt aaatggtgat catgtaaccc atcctgactt catgttgtat
     2581 gacgctcttg atgttgtttt atacatggac ccaatgtgcc tggatgcgtt cccaaaatta
     2641 gtttgtttta aaaaacgtat tgaagctatc ccacaaattg ataagtactt gaaatccagc
     2701 aagtatatag catggccttt gcagggctgg caagccacgt ttggtggtgg cgaccatcct
     2761 ccaaaatcgg atctggaagt tctgttccag gggcccctgg gatccatggc tcggacaaac
     2821 cagtacccga gacagccggc tggagggaag cgaagaaagg gtgcaactcc gaagagaagg
     2881 gaaccgatcg agcagcagaa tcgaccgccg cgtccaccgt ccccgcgacc cgtggtgtca
     2941 cccgcgagct taccgtcaag cggtccgctc agtcggaaat caccgggtgg cagaaatcca
     3001 cctcgagctt aagaattccc gggtcgactc gagcggccgc atcgtgactg actgacgatc
     3061 tgcctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg
     3121 gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg
     3181 ggtgttggcg ggtgtcgggg cgcagccatg acccagtcac gtagcgatag cggagtgtat
     3241 aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3301 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3361 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3421 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     3481 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     3541 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     3601 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     3661 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     3721 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     3781 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3841 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3901 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3961 ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa
     4021 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4081 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4141 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4201 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4261 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4321 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4381 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4441 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     4501 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     4561 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     4621 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     4681 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     4741 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     4801 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4861 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4921 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4981 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5041 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5101 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5161 gataaccgta ttaccgcctt tgagtgag
//
