LOCUS       sequence-343276-        6525 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription template for rat CENP-C (N-ter EGFP tagged).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6525)
  AUTHORS   Kumon T, Ma J, Akins RB, Stefanik D, Nordgren CE, Kim J, Levine MT, 
            Lampson MA
  TITLE     Parallel pathways for recruiting effector proteins determine 
            centromere drive and suppression.
  JOURNAL   Cell. 2021 Sep 16;184(19):4904-4918.e11. doi: 
            10.1016/j.cell.2021.07.037. Epub 2021 Aug 24.
  PUBMED    34433012
REFERENCE   2  (bases 1 to 6525)
  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.cell.2021.07.037"; journalName: "Cell"; date: 
            "2021-09-16- 16"; volume: "184"; issue: "19"; pages: "4904-4918.e11"
COMMENT     Sequence Label: pIVT-EGFP-ratCENP-C
FEATURES             Location/Qualifiers
     source          1..6525
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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     complement(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2246..2264
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2246..2263
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2276..2295
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2276..2294
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     5'UTR           2337..2379
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     regulatory      2399..2408
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2405..2407,2408..2410,2411..3121)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2405..2407,2408..2410,2411..3121)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2450..2471)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2711..2730)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3058..3079
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     3'UTR           5943..6068
                     /label=Xenopus globin 3'-UTR
                     /note="3'-UTR of the major beta-globin gene of Xenopus 
                     laevis"
     primer_bind     complement(6131..6148)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6131..6147)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6140..6162)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6356..6375)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6475..6497
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctc caacgattta ggtgacacta tagaagagaa ggaattaata
     2281 cgactcacta tagggagaga gagagaatta ccctcactaa agggaggaga agcttgcttg
     2341 ttctttttgc agaagctcag aataaacgct caactttggc agatcaagct tgcatgccgc
     2401 caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct
     2461 ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac
     2521 ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc
     2581 caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat
     2641 gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat
     2701 cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac
     2761 cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg
     2821 gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa
     2881 gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct
     2941 cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa
     3001 ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat
     3061 ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa
     3121 ggtcgactct agaggatccc cgggtaccat ggcctcgttc catctggatc atctaaagaa
     3181 ctaccacaga agattttgcc ggccttccag agcacccaac attcctacaa agcaaggcca
     3241 gaacatgctg gaaatcttac aagattgttt tgaagaacag aattctacag agtcattgtc
     3301 ttgttcaaca cctaaagaga aagacattta tgttcagtca acaggcaaag agcgccaagc
     3361 atcacattca gaatcagttc cgatttcttc aaggaggaag gaagcttctt tgcaagttcc
     3421 tgcaaaacca agtgaccctg ctgatgaacc agttcaggct aatgaagttc atcagaaaat
     3481 tctgtctact gatgtggtct ccaaaagtac accagactca accaaaacat caagtaaagg
     3541 agtaaaaggt caccatggtg cgtctgatga attgtactta cctggtggct cccctgtggt
     3601 tcttctgggt gcaagagtgt ccgggtcaca gaaagctatt tcatctggtg gccaaaaaag
     3661 agtgacttca aggcgttcag cagatatgca gtcttcaaat acagacattt cttttaaaac
     3721 caggaaaagg ctaaacttcg aagataaagt tatttcgaac acagcagaaa tagagagcag
     3781 tgcatcacaa gtagaggata gcatatccga ggaacaagaa gggacatcat cagaaacttc
     3841 tcagaagaga gacgatctga gctctgaaat tcaaccacaa tctaaaaaga gcttttcaga
     3901 gttgtattta gaaactgtca acaggaaaag taaatgcagt tctgttgtta ggcacacagc
     3961 agcagttcca tctcctccgt atcctccgaa tgatatgaaa ttattagaag atgaatttat
     4021 tattgatggg ccagatagaa gcttttcaag tcaaccttgg gtggtgatac caaggaaagg
     4081 ccaccacatg ccatttcctg aaaacattgc agtgcctcaa ggtaagaagt caagagagaa
     4141 acctcatcgt ctgtccgaga agactttggt aagtaacaca cagactgata agactggccc
     4201 agtagaggaa gctcagctct ctgttgagga aaaactggga actactttga caaatgagtt
     4261 ggaaaatgat tgtagatcta cagaaaataa aacgcactct gagaatgcga aaaaaccatc
     4321 tgcacggaaa aggactgtga aacagaagca gagaagacca tctaagccta acatagctca
     4381 agagctcagt atgggacaga atgaaactga aaataggaat atgtcaaaaa ttggccaaga
     4441 caagttacaa ataaattcaa aaagaaatat ggaagaccat gaagaggtga ggaacgagcc
     4501 tacccccaag aaacctgtgc ctgctcttgg aaataagaaa gaaaaggaca gcacccaagc
     4561 taataaggaa aaatctagaa agaaatgttt ttctagagag tccaagagga aatctgttcc
     4621 caaagaagta actttggctt ccaggagagg tcgtagaact tctcagcatc cgtctgagtg
     4681 gtggttggta aaaccaagtg aaggttctgt tgatggaaat tcttcaaagg agaatgaatc
     4741 atcaatttta tatccaaata gaaaaaaaca gactaagaga aatcacgtat ctgagaatac
     4801 tgagaagaaa cctgttccat cgaaaagaaa gaagaaagag atcagctcaa gagaactgaa
     4861 gtctttaaat gtaaagggtt ctggaggaaa tgatgatatt tctacttccc agaggaagcc
     4921 gttgcaaaac attgacgcag atccaactca gaagagcctt cattgttctg gacctaccag
     4981 aggctctgaa gatcaaaaca gtgatatgat atcacagaat gttcatttaa agactcgtag
     5041 tgagcaacat agaagcaaga cacaaatgga atctacttca aattcggaag tgtgtaaaca
     5101 ttcagtgtgg gaagaaagtg gaccttccag gttcaagaat cttgaaatgc ctggacgcag
     5161 taattctgaa gtgggtgacg aaaaggacca gaagagtttg gatttgaaaa caagaagttc
     5221 taatatggta ccgaatagga atttacacca caaattagta ttgccctcca actcaccaaa
     5281 tgttcgtaga tctaatagaa tacgtttgaa acctctggaa tattggcgag gggaacgaat
     5341 agattatcaa gaaagctcat caggaagact tgtgcttgaa atagtgtccc cagcttcaga
     5401 atcagtgaaa agaaaggcca aaagaaatct ggacaaagtc aacaaagaaa ctaacaagaa
     5461 acggatccat cttgataacc ctacaaagac taagataaag gtgagtctgg atattcctct
     5521 gggagatccc ttccaggcaa cattggcaaa ggacccggaa acaagagaga ttgtccccat
     5581 ggatcttata agaccacgag atacatacca cttttttgtt gagcaacatg gtttgaaagt
     5641 ttataaaaca ttgaatacaa catttttttc tactggaaaa ttggttttag gaccccatga
     5701 agaaaaagga aaacagcatg ttggtcaaga tatattagtt ttttatgtta actttggtga
     5761 ccttctatgt acattacatg aaacacctta tatgctaacg accggggatt cattctatgt
     5821 tccttcaggt aatcattaca acattaaaaa cctcttgaat gtggaaagtt gtcttctttt
     5881 tactcaaata aaaaggtgag aattcgccct tctagtgact gactaggatc tggttaccac
     5941 taaaccagcc tcaagaacac ccgaatggag tctctaagct acataatacc aacttacact
     6001 ttacaaaatg ttgtccccca aaatgtagcc attcgtatct gctcctaata aaaaraaagt
     6061 ttcttcacat tctaaaaaaa aaaaaaaaaa aaaaaaaaaa aaaanncccc ccccccccaa
     6121 gggcgaattc actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     6181 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     6241 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatggcgc ctgatgcggt
     6301 attttctcct tacgcatctg tgcggtattt cacaccgcat atggtgcact ctcagtacaa
     6361 tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc gctgacgcgc
     6421 cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc gtctccggga
     6481 gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcga
//
