LOCUS       sequence-221625-        7127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of exogenous LacI-GFP-HEC1 delta 207.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7127)
  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 7127)
  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-d207
FEATURES             Location/Qualifiers
     source          1..7127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        135..438
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        439..642
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     593..613
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     complement(693..712)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             1754..1774
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     regulatory      1793..1802
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1799..1801,1802..1804,1805..2515)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1799..1801,1802..1804,1805..2515)
                     /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(1844..1865)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2105..2124)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2452..2473
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3989..4110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4026..4045)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4080..4099
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4117..4572)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4254..4275)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4466..4485
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4599..4703
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4705..5062
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4705..4896
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4728..4748)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4913..5048
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5097..5891
                     /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             5097..5891
                     /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(5151..5170)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5761..5780
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6079..6098)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6123..6170
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6499..7087
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6988..7007
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
       61 ttaccgccat gcattagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      121 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      181 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      241 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
      301 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      361 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      421 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      481 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      541 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      601 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
      661 atccgctagc atggtgaaac cagtaacgtt atacgatgtc gcagagtatg ccggtgtctc
      721 ttatcagacc gtttcccgcg tggtgaacca ggccagccac gtttctgcga aaacgcggga
      781 aaaagtggaa gcggcgatgg cggagctgaa ttacattccc aaccgcgtgg cacaacaact
      841 ggcgggcaaa cagtcgttgc tgattggcgt tgccacctcc agtctggccc tgcacgcgcc
      901 gtcgcaaatt gtcgcggcga ttaaatctcg cgccgatcaa ctgggtgcca gcgtggtggt
      961 gtcgatggta gaacgaagcg gcgtcgaagc ctgtaaagcg gcggtgcaca atcttctcgc
     1021 gcaacgcgtc agtgggctga tcattaacta tccgctggat gaccaggatg ccattgctgt
     1081 ggaagctgcc tgcactaatg ttccggcgtt atttcttgat gtctctgacc agacacccat
     1141 caacagtatt attttctccc atgaagacgg tacgcgactg ggcgtggagc atctggtcgc
     1201 attgggtcac cagcaaatcg cgctgttagc gggcccatta agttctgtct cggcgcgtct
     1261 gcgtctggct ggctggcata aatatctcac tcgcaatcaa attcagccga tagcggaacg
     1321 ggaaggcgac tggagtgcca tgtccggttt tcaacaaacc atgcaaatgc tgaatgaggg
     1381 catcgttccc actgcgatgc tggttgccaa cgatcagatg gcgctgggcg caatgcgcgc
     1441 cattaccgag tccgggctgc gcgttggtgc ggatatctcg gtagtgggat acgacgatac
     1501 cgaagacagc tcatgttata tcccgccgtt aaccaccatc aaacaggatt ttcgcctgct
     1561 ggggcaaacc agcgtggacc gcttgctgca actctctcag ggccaggcgg tgaagggcaa
     1621 tcagctgttg cccgtctcac tggtgaaaag aaaaaccacc ctggcgccca atacgcaaac
     1681 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgaag
     1741 cagcctgagg cctcctaaga agaagaggaa ggttgggcta gcgctaccgg tcgccaccat
     1801 ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1861 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1921 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1981 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     2041 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     2101 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2161 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2221 gctggagtac aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg
     2281 catcaaggtg aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2341 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2401 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2461 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtccgg
     2521 actcagatcc gggcagcctt ggggagaaga aactgaagat ggaattatgc ataataagtt
     2581 gtttttggac tacaccataa aatgctatga gagttttatg agtggtgccg acagctttga
     2641 tgagatgaat gcagagctgc agtcaaaact gaaggattta tttaatgtgg atgcttttaa
     2701 gctggaatca ttagaagcaa aaaacagagc attgaatgaa cagattgcaa gattggaaca
     2761 agaaagagaa aaagaaccga atcgtctaga gtcgttgaga aaactgaagg cttccttaca
     2821 aggagatgtt caaaagtatc aggcatacat gagcaatttg gagtctcatt cagccattct
     2881 tgaccagaaa ttaaatggtc tcaatgagga aattgctaga gtagaactag aatgtgaaac
     2941 aataaaacag gagaacactc gactacagaa tatcattgac aaccagaagt actcagttgc
     3001 agacattgag cgaataaatc atgaaagaaa tgaattgcag cagactatta ataaattaac
     3061 caaggacctg gaagctgaac aacagaagtt gtggaatgag gagttaaaat atgccagagg
     3121 caaagaagcg attgaaacac aattagcaga gtatcacaaa ttggctagaa aattaaaact
     3181 tattcctaaa ggtgctgaga attccaaagg ttatgacttt gaaattaagt ttaatcccga
     3241 ggctggtgcc aactgccttg tcaaatacag ggctcaagtt tatgtacctc ttaaggaact
     3301 cctgaatgaa actgaagaag aaattaataa agccctaaat aaaaaaatgg gtttggagga
     3361 tactttagaa caattgaatg caatgataac agaaagcaag agaagtgtga gaactctgaa
     3421 agaagaagtt caaaagctgg atgatcttta ccaacaaaaa attaaggaag cagaggaaga
     3481 ggatgaaaaa tgtgccagtg agcttgagtc cttggagaaa cacaagcacc tgctagaaag
     3541 tactgttaac caggggctca gtgaagctat gaatgaatta gatgctgttc agcgggaata
     3601 ccaactagtt gtgcaaacca cgactgaaga aagacgaaaa gtgggaaata acttgcaacg
     3661 tctgttagag atggttgcta cacatgttgg gtctgtagag aaacatcttg aggagcagat
     3721 tgctaaagtt gatagagaat atgaagaatg catgtcagaa gatctctcgg aaaatattaa
     3781 agagattaga gataagtatg agaagaaagc tactctaatt aagtcttctg aagaatgact
     3841 cgacggtacc gcgggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     3901 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3961 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     4021 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     4081 tggtttgtcc aaactcatca atgtatctta acgcgtaaat tgtaagcgtt aatattttgt
     4141 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     4201 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     4261 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4321 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4381 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4441 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4501 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4561 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4621 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4681 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4741 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4801 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4861 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4921 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4981 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     5041 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     5101 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     5161 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     5221 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     5281 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     5341 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     5401 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     5461 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     5521 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     5581 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5641 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5701 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5761 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5821 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5881 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5941 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     6001 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     6061 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     6121 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     6181 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     6241 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     6301 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     6361 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     6421 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     6481 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     6541 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6601 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     6661 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6721 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6781 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6841 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6901 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6961 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     7021 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     7081 atggaaaaac gccagcaacg cggccttttt acggttcctg gcctttt
//
