LOCUS       sequence-205246-        5578 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expression of vsv-CenpB 1-166.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5578)
  AUTHORS   Liu D, Vader G, Vromans MJ, Lampson MA, Lens SM
  TITLE     Sensing chromosome bi-orientation by spatial separation of aurora B 
            kinase from kinetochore substrates.
  JOURNAL   Science. 2009 Mar 6;323(5919):1350-3. doi: 10.1126/science.1167000. 
            Epub 2009 Jan 15.
  PUBMED    19150808
REFERENCE   2  (bases 1 to 5578)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.1167000"; journalName: "Science"; date: "2009-03-6-
            6"; volume: "323"; issue: "5919"; pages: "1350-3"
COMMENT     Sequence Label: pCR3-vsv-CenpB 1-166
FEATURES             Location/Qualifiers
     source          1..5578
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        10..389
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        390..593
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     544..564
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     638..657
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        638..656
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             691..723
                     /codon_start=1
                     /product="epitope tag from vesicular stomatitis virus G 
                     protein"
                     /label=VSV-G tag
                     /translation="YTDIEMNRLGK"
     promoter        complement(1293..1311)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1294..1311)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1331..1348)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1337..1561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(1693..2281)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1773..1792)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    2610..2657
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     2682..2701
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(2889..3677)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDQEHQGL
                     APAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALA
                     TRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3000..3019)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     3604..3623
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(3712..4069)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3726..3861
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        3878..4069
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4026..4046
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4129..4920,4921..4989))
                     /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     4752..4771
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4990..5094)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5121..5576
                     /direction=RIGHT
                     /label=M13 ori
                     /note="M13 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5208..5227)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5418..5439
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
       61 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      121 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      181 caatagggac tttccattga cgtcaatggg tggactattt acggtaaact gcccacttgg
      241 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      301 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      361 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      421 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      481 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      541 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
      601 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
      661 cccaagcttg gtaccggatc cgccaccatg tacaccgaca tcgagatgaa ccggttgggc
      721 aaggaattcg gccccaagag gcgacagctg acgttccggg agaagtcacg gatcatccag
      781 gaggtggagg agaatccgga cctgcgcaag ggcgagatcg cgcggcgctt caacatcccg
      841 ccgtccacgc tgagcacgat cctgaagaac aagcgcgcca tcctggcgtc ggagcgcaag
      901 tacggggtgg cctccacctg ccgcaagacc aacaagctgt ctccctacga caagctcgag
      961 ggcttgctca tcgcctggtt ccagcagatc cgcgccgccg gcctgccggt caagggcatc
     1021 atcctcaagg agaaggcgct gcgcatagcc gaggagctgg gcatggacga cttcaccgcc
     1081 tccaacggct ggctggaccg cttccgccgg cgccacggcg tggtgtcctg cagcggcgtg
     1141 gcccgcgccc gcgcgcgaaa cgctgccccc cgcaccccgg cggcgcctgc cagtccggcc
     1201 gcggtgccct cggagggcgg gagcactgaa ttctgcagat atccatcaca ctggcggccg
     1261 ctcgagtaga tgactagtct agagggccct attctatagt gtcacctaaa tgctagagct
     1321 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1381 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1441 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     1501 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     1561 gcttctgagg cggaaagaac cagtggcggt aatacggtta tccacagaat caggggataa
     1621 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     1681 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     1741 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     1801 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     1861 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     1921 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     1981 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     2041 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     2101 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     2161 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     2221 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     2281 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     2341 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     2401 atgaagtttt aaatcaatct aaagtatata tgagtaacct gaggctatgg cagggcctgc
     2461 cgccccgacg ttggctgcga gccctgggcc ttcacccgaa cttggggggt ggggtgggga
     2521 aaaggaagaa acgcgggcgt attggcccca atggggtctc ggtggggtat cgacagagtg
     2581 ccagccctgg gaccgaaccc cgcgtttatg aacaaacgac ccaacaccgt gcgttttatt
     2641 ctgtcttttt attgccgtca tagcgcgggt tccttccggt attgtctcct tccgtgtttc
     2701 agttagcctc cccctagggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     2761 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     2821 ggaatcgaaa tctcgtgatg gcaggttggg cgtcgcttgg tcggtcattt cgaaccccag
     2881 agtcccgctc agaagaactc gtcaagaagg cgatagaagg cgatgcgctg cgaatcggga
     2941 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
     3001 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
     3061 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
     3121 tgggtcacga cgagatcctc gccgtcgggc atgctcgcct tgagcctggc gaacagttcg
     3181 gctggcgcga gcccctgatg ctcttgatca tcctgatcga caagaccggc ttccatccga
     3241 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca
     3301 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg
     3361 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct
     3421 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc
     3481 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga
     3541 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt
     3601 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat
     3661 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa
     3721 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgaggcggc
     3781 ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat gggcggaact
     3841 gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg ctgactaatt
     3901 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacacctggt
     3961 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     4021 tccacaccct aactgacaca cattccacag ctggttcttt ccgcctcagg actcttcctt
     4081 tttcaataaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     4141 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     4201 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     4261 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     4321 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     4381 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     4441 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     4501 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     4561 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     4621 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     4681 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     4741 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     4801 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     4861 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     4921 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     4981 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     5041 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     5101 tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     5161 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     5221 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     5281 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     5341 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     5401 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     5461 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     5521 gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta caatttac
//
