LOCUS       sequence-200660-        5330 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription, zf centrin N-terminally tagged with mkate2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5330)
  AUTHORS   Strzyz PJ, Lee HO, Sidhaye J, Weber IP, Leung LC, Norden C
  TITLE     Interkinetic nuclear migration is centrosome independent and ensures
            apical cell division to maintain tissue integrity.
  JOURNAL   Dev Cell. 2015 Jan 26;32(2):203-19. doi: 
            10.1016/j.devcel.2014.12.001. Epub 2015 Jan 15.
  PUBMED    25600237
REFERENCE   2  (bases 1 to 5330)
  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.2014.12.001"; journalName: "Dev Cell"; date: 
            "2015-01-26- 26"; volume: "32"; issue: "2"; pages: "203-19"
COMMENT     Sequence Label: pCS2+ centrin-mKate2
FEATURES             Location/Qualifiers
     source          1..5330
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             106..801
                     /codon_start=1
                     /product="monomeric far-red fluorescent protein (Shcherbo 
                     et al., 2009)"
                     /label=mKate2
                     /note="mammalian codon-optimized"
                     /translation="MVSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMRIK
                     AVEGGPLPFAFDILATSFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLT
                     ATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRADMAL
                     KLVGGGHLICNLKTTYRSKKPAKNLKMPGVYYVDRRLERIKEADKETYVEQHEVAVARY
                     CDLPSKLGHR"
     polyA_signal    1379..1513
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1403..1422)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1457..1476
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1626..1646)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1626..1644)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1665..1681)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1665..1681)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1678..1700)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1689..1705
                     /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)."
     promoter        complement(join(1713..1719,1720..1737,1738..1743))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1758..1779
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1896..1913)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2067..2655)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2147..2166)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2826..3617,3618..3686))
                     /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     3449..3468
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3687..3791)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3817..4272)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3954..3975)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4166..4185
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4346..5330
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atcccatcga ttcgaattcc catggatggt gagcgagctg
      121 attaaggaga acatgcacat gaagctgtac atggagggca ccgtgaacaa ccaccacttc
      181 aagtgcacat ccgagggcga aggcaagccc tacgagggca cccagaccat gagaatcaag
      241 gcggtcgagg gcggccctct ccccttcgcc ttcgacatcc tggctaccag cttcatgtac
      301 ggcagcaaaa ccttcatcaa ccacacccag ggcatccccg acttctttaa gcagtccttc
      361 cccgagggct tcacatggga gagagtcacc acatacgaag acgggggcgt gctgaccgct
      421 acccaggaca ccagcctcca ggacggctgc ctcatctaca acgtcaagat cagaggggtg
      481 aacttcccat ccaacggccc tgtgatgcag aagaaaacac tcggctggga ggcctccacc
      541 gagaccctgt accccgctga cggcggcctg gaaggcagag ccgacatggc cctgaagctc
      601 gtgggcgggg gccacctgat ctgcaacttg aagaccacat acagatccaa gaaacccgct
      661 aagaacctca agatgcccgg cgtctactat gtggacagaa gactggaaag aatcaaggag
      721 gccgacaaag agacctacgt cgagcagcac gaggtggctg tggccagata ctgcgacctc
      781 cctagcaaac tggggcacag agatatcatg gcgtccggct tcaggaaaag cagtgcttca
      841 gcaaaccaga ggaagaaagc aggacccaaa ccagagctga cagaagagca gaagcaggag
      901 atcaaagagg cgttcgatct gttcgacaca gacggctctg gaaccatcga tgtcaaggaa
      961 ctgaaggttg cgatgcgagc tctggggttt gagccgaaga aagaagagat aaagaagatg
     1021 atcgctgata ttgataaaga aggatcgggc gtaatcggct tcagcgattt cctttctatg
     1081 atgacacaga aaatgagtga gaaagactca aaagaagaga tcctgaaggc tttccggctg
     1141 tttgacgatg actgcactgg gaaaatctcc ttcaagaacc tgaagcgagt ggctaaagag
     1201 ctcggagaga acctgacgga tgaggagctg caggaaatga tcgacgaggc cgacagagac
     1261 ggtgacgggg agattaatga gcaggagttt ctgcggatta tgaagaaaac caatctgtac
     1321 tgaagatctg ctagcaggcc tctcgagcct ctagaactat agtgagtcgt attacgtaga
     1381 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1441 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1501 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1561 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     1621 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     1681 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     1741 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1801 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1861 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1921 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1981 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2041 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2101 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2161 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2221 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2281 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2341 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2401 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2461 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     2521 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2581 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2641 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2701 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2761 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2821 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2881 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2941 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3001 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3061 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3121 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3181 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3241 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3301 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3361 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3421 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3481 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3541 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3601 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3661 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3721 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3781 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     3841 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3901 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3961 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     4021 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     4081 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     4141 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     4201 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     4261 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4321 gggcctcttc gctattacgc cagtcgacca tagccaattc aatatggcgt atatggactc
     4381 atgccaattc aatatggtgg atctggacct gtgccaattc aatatggcgt atatggactc
     4441 gtgccaattc aatatggtgg atctggaccc cagccaattc aatatggcgg acttggcacc
     4501 atgccaattc aatatggcgg acttggcact gtgccaactg gggaggggtc tacttggcac
     4561 ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt ccgccatatt gaattggcat
     4621 ggtgccaata atggcggcca tattggctat atgccaggat caatatatag gcaatatcca
     4681 atatggccct atgccaatat ggctattggc caggttcaat actatgtatt ggccctatgc
     4741 catatagtat tccatatatg ggttttccta ttgacgtaga tagcccctcc caatgggcgg
     4801 tcccatatac catatatggg gcttcctaat accgcccata gccactcccc cattgacgtc
     4861 aatggtctct atatatggtc tttcctattg acgtcatatg ggcggtccta ttgacgtata
     4921 tggcgcctcc cccattgacg tcaattacgg taaatggccc gcctggctca atgcccattg
     4981 acgtcaatag gaccacccac cattgacgtc aatgggatgg ctcattgccc attcatatcc
     5041 gttctcacgc cccctattga cgtcaatgac ggtaaatggc ccacttggca gtacatcaat
     5101 atctattaat agtaacttgg caagtacatt actattggaa ggacgccagg gtacattggc
     5161 agtactccca ttgacgtcaa tggcggtaaa tggcccgcga tggctgccaa gtacatcccc
     5221 attgacgtca atggggaggg gcaatgacgc aaatgggcgt tccattgacg taaatgggcg
     5281 gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg tttagggaac
//
