LOCUS       sequence-221624-        7748 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of exogenous LacI-GFP-HEC1 WT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7748)
  AUTHORS   Nijenhuis W, von Castelmur E, Littler D, De Marco V, Tromer E, 
            Vleugel M, van Osch MH, Snel B, Perrakis A, Kops GJ
  TITLE     A TPR domain-containing N-terminal module of MPS1 is required for 
            its kinetochore localization by Aurora B.
  JOURNAL   J Cell Biol. 2013 Apr 15;201(2):217-31. doi: 10.1083/jcb.201210033. 
            Epub 2013 Apr 8.
  PUBMED    23569217
REFERENCE   2  (bases 1 to 7748)
  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: 
            "10.1083/jcb.201210033"; journalName: "J Cell Biol"; date: 
            "2013-04-15- 15"; volume: "201"; issue: "2"; pages: "217-31"
COMMENT     Sequence Label: pLacI-GFP-HEC1 WT
FEATURES             Location/Qualifiers
     source          1..7748
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        56..359
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        360..563
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     514..534
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(614..633)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1675..1695
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1714..1723
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1720..1722,1723..1725,1726..2436)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1720..1722,1723..1725,1726..2436)
                     /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(1765..1786)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2026..2045)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2373..2394
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4531..4652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4568..4587)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4622..4641
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4659..5114)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4796..4817)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5008..5027
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5141..5245
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5247..5604
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5247..5438
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5270..5290)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5455..5590
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5639..6433
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5639..6433
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5693..5712)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6303..6322
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6621..6640)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6665..6712
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7041..7629
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7530..7549
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
       61 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      121 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      181 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      241 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      301 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      361 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      421 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      481 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      541 gggaggtcta tataagcaga gctggtttag tgaaccgtca gatccgctag catggtgaaa
      601 ccagtaacgt tatacgatgt cgcagagtat gccggtgtct cttatcagac cgtttcccgc
      661 gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg aaaaagtgga agcggcgatg
      721 gcggagctga attacattcc caaccgcgtg gcacaacaac tggcgggcaa acagtcgttg
      781 ctgattggcg ttgccacctc cagtctggcc ctgcacgcgc cgtcgcaaat tgtcgcggcg
      841 attaaatctc gcgccgatca actgggtgcc agcgtggtgg tgtcgatggt agaacgaagc
      901 ggcgtcgaag cctgtaaagc ggcggtgcac aatcttctcg cgcaacgcgt cagtgggctg
      961 atcattaact atccgctgga tgaccaggat gccattgctg tggaagctgc ctgcactaat
     1021 gttccggcgt tatttcttga tgtctctgac cagacaccca tcaacagtat tattttctcc
     1081 catgaagacg gtacgcgact gggcgtggag catctggtcg cattgggtca ccagcaaatc
     1141 gcgctgttag cgggcccatt aagttctgtc tcggcgcgtc tgcgtctggc tggctggcat
     1201 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
     1261 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
     1321 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
     1381 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
     1441 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
     1501 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
     1561 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
     1621 gccgattcat taatgcagct ggcacgacag gtttcccgaa gcagcctgag gcctcctaag
     1681 aagaagagga aggttgggct agcgctaccg gtcgccacca tggtgagcaa gggcgaggag
     1741 ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag
     1801 ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc
     1861 atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac
     1921 ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc
     1981 gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac
     2041 aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag
     2101 ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac
     2161 agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag
     2221 atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc
     2281 cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc
     2341 ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc
     2401 gccgggatca ctctcggcat ggacgagctg tacaagtccg gactcagatc catgaagcgc
     2461 agttcagttt ccagcggtgg tgctggccgc ctctccatgc aggagttaag atcccaggat
     2521 gtaaataaac aaggcctcta tacccctcaa accaaagaga aaccaacctt tggaaagttg
     2581 agtataaaca aaccgacatc tgaaagaaaa gtctcgctat ttggcaaaag aactagtgga
     2641 catggatcac ggaatagtca acttggtata ttttccagtt ctgagaaaat caaggacccg
     2701 agaccactta acgacaaagc attcattcag cagtgtattc gacaactctg tgagtttctt
     2761 acagaaaatg gttatgcaca taatgtgtcc atgaaatctc tacaagctcc ctctgttaaa
     2821 gacttcctga agatcttcac atttctttat ggcttcctgt gcccctcata cgaacttcct
     2881 gacacaaagt ttgaagaaga ggttccaaga atctttaaag accttgggta tccttttgca
     2941 ctatccaaaa gctccatgta cacagtgggg gctcctcata catggcctca cattgtggca
     3001 gccttagttt ggctaataga ctgcatcaag atacatactg ccatgaaaga aagctcacct
     3061 ttatttgatg atgggcagcc ttggggagaa gaaactgaag atggaattat gcataataag
     3121 ttgtttttgg actacaccat aaaatgctat gagagtttta tgagtggtgc cgacagcttt
     3181 gatgagatga atgcagagct gcagtcaaaa ctgaaggatt tatttaatgt ggatgctttt
     3241 aagctggaat cattagaagc aaaaaacaga gcattgaatg aacagattgc aagattggaa
     3301 caagaaagag aaaaagaacc gaatcgtcta gagtcgttga gaaaactgaa ggcttcctta
     3361 caaggagatg ttcaaaagta tcaggcatac atgagcaatt tggagtctca ttcagccatt
     3421 cttgaccaga aattaaatgg tctcaatgag gaaattgcta gagtagaact agaatgtgaa
     3481 acaataaaac aggagaacac tcgactacag aatatcattg acaaccagaa gtactcagtt
     3541 gcagacattg agcgaataaa tcatgaaaga aatgaattgc agcagactat taataaatta
     3601 accaaggacc tggaagctga acaacagaag ttgtggaatg aggagttaaa atatgccaga
     3661 ggcaaagaag cgattgaaac acaattagca gagtatcaca aattggctag aaaattaaaa
     3721 cttattccta aaggtgctga gaattccaaa ggttatgact ttgaaattaa gtttaatccc
     3781 gaggctggtg ccaactgcct tgtcaaatac agggctcaag tttatgtacc tcttaaggaa
     3841 ctcctgaatg aaactgaaga agaaattaat aaagccctaa ataaaaaaat gggtttggag
     3901 gatactttag aacaattgaa tgcaatgata acagaaagca agagaagtgt gagaactctg
     3961 aaagaagaag ttcaaaagct ggatgatctt taccaacaaa aaattaagga agcagaggaa
     4021 gaggatgaaa aatgtgccag tgagcttgag tccttggaga aacacaagca cctgctagaa
     4081 agtactgtta accaggggct cagtgaagct atgaatgaat tagatgctgt tcagcgggaa
     4141 taccaactag ttgtgcaaac cacgactgaa gaaagacgaa aagtgggaaa taacttgcaa
     4201 cgtctgttag agatggttgc tacacatgtt gggtctgtag agaaacatct tgaggagcag
     4261 attgctaaag ttgatagaga atatgaagaa tgcatgtcag aagatctctc ggaaaatatt
     4321 aaagagatta gagataagta tgagaagaaa gctactctaa ttaagtcttc tgaagaatga
     4381 ctcgacggta ccgcgggccc gggatccacc ggatctagat aactgatcat aatcagccat
     4441 accacatttg tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg
     4501 aaacataaaa tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac
     4561 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4621 tgtggtttgt ccaaactcat caatgtatct taacgcgtaa attgtaagcg ttaatatttt
     4681 gttaaaattc gcgttaaatt tttgttaaat cagctcattt tttaaccaat aggccgaaat
     4741 cggcaaaatc ccttataaat caaaagaata gaccgagata gggttgagtg ttgttccagt
     4801 ttggaacaag agtccactat taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt
     4861 ctatcagggc gatggcccac tacgtgaacc atcaccctaa tcaagttttt tggggtcgag
     4921 gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc cgatttagag cttgacgggg
     4981 aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc
     5041 gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc
     5101 gctacagggc gcgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5161 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5221 tcaataatat tgaaaaagga agagtcctga ggcggaaaga accagctgtg gaatgtgtgt
     5281 cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     5341 ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     5401 caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg
     5461 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     5521 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     5581 tttggaggcc taggcttttg caaagatcga tcaagagaca ggatgaggat cgtttcgcat
     5641 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     5701 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     5761 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     5821 agacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     5881 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     5941 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     6001 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     6061 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     6121 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg
     6181 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     6241 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     6301 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     6361 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     6421 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     6481 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6541 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6601 caccctaggg ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct
     6661 atgacggcaa taaaaagaca gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc
     6721 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     6781 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     6841 ctcgcagcca acgtcggggc ggcaggccct gccatagcct caggttactc atatatactt
     6901 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6961 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     7021 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     7081 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     7141 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     7201 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     7261 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     7321 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     7381 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     7441 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     7501 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     7561 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7621 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7681 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccatg
     7741 cattagtt
//
