LOCUS       sequence-200654-        8440 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  in vitro transcription.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8440)
  AUTHORS   Weber IP, Ramos AP, Strzyz PJ, Leung LC, Young S, Norden C
  TITLE     Mitotic position and morphology of committed precursor cells in the 
            zebrafish retina adapt to architectural changes upon tissue 
            maturation.
  JOURNAL   Cell Rep. 2014 Apr 24;7(2):386-97. doi: 
            10.1016/j.celrep.2014.03.014. Epub 2014 Apr 3.
  PUBMED    24703843
REFERENCE   2  (bases 1 to 8440)
  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.celrep.2014.03.014"; journalName: "Cell Rep"; date: 
            "2014-04-24- 24"; volume: "7"; issue: "2"; pages: "386-97"
COMMENT     Sequence Label: pCS2+-rasmKate
FEATURES             Location/Qualifiers
     source          1..8440
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(34..462)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(144..165)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     356..375
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     589..611
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     603..620
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     604..620
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     630..649
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        630..648
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    join(778..789,790..952)
                     /label=Tol2 right end
                     /note="right end of the Tol2 transposon from the medaka 
                     fish Oryzias latipes (Kawakami, 2007)"
     protein_bind    1324..1344
                     /gene="mutant version of attB"
                     /label=attB4
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     protein_bind    2864..2888
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2891..3586
                     /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"
     protein_bind    complement(3666..3690)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    3695..3829
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3719..3738)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3773..3792
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     misc_feature    join(5970..6157,6158..6169)
                     /label=Tol2 left end
                     /note="left end of the Tol2 transposon from the medaka fish
                     Oryzias latipes (Kawakami, 2007)"
     primer_bind     complement(6203..6219)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6203..6219)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(6256..6276)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6256..6274)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6295..6311)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6295..6311)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6308..6330)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6319..6335
                     /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(6343..6349,6350..6367,6368..6373))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6388..6409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6526..6543)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6697..7285)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6777..6796)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7456..8247,8248..8316))
                     /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     8079..8098
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8317..8421)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ccacctaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
       61 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
      121 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
      181 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
      241 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
      301 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
      361 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
      421 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc
      481 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
      541 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
      601 gttgtaaaac gacggccagt gagcgcgcgt aatacgactc actatagggc gaattgggta
      661 caggtcattg cacaacacca gaaatgccct ctgatctgca aaagacgtga atatctgttc
      721 agacacccat atccactctg ttccacacag gtcagaggtt tgtccaggag ttcttgacag
      781 aggtgtaaaa agtactcaaa aattttactc aagtgaaagt acaagtactt agggaaaatt
      841 ttactcaatt aaaagtaaaa gtatctggct agaatcttac ttgagtaaaa gtaaaaaagt
      901 actccattaa aattgtactt gagtattaag gaagtaaaag taaaagcaag aaagaaaact
      961 agagattctt gtttaagctt ttaatctcaa aaaacattaa atgaaatgca tacaaggttt
     1021 tatcctgctt tagaactgtt tgtatttaat tatcaaacta taagacagac aatctaatgc
     1081 cagtacacgc tactcaaagt tgtaaaacct cagatttaac ttcagtagaa gctgattctc
     1141 aaaattgtta gtgtcaagcc tagctctttt ggggctgaaa agcaatcctg cagtgctgaa
     1201 aagcctctca caggcagccg atgcgggaag aggtgtatta gtcttgatag agaggctgca
     1261 aatagcagga aacgtgagca gagactccct ggtgtctgaa acacaggcca gatgggccga
     1321 tatcaacttt gtatagaaaa gttgcttcag gggtgtcgct tggttatttc caaaaatcaa
     1381 attaatttta ttaaactatt agaacgagca tgttttgtct atatgctaca gaagataaaa
     1441 aataatagga gttaacagtt ataaaacaac acactttgtt tctattgatt gttgaccaca
     1501 ctggggtctc attaagttag attaaagaca cactaactgg gtcaaaagca gcagattgat
     1561 ttcatagcac cagggtaaac tttctaacac ttttacggca atcatataca ttaaaattaa
     1621 atacagacca cgactgaaca aggaggatga tctccaatat taaacaaaga gacttgtgcc
     1681 tatttctctg agggtaaaca tgacctctca agttagcaag ttggttttaa cactacaaaa
     1741 atagttaaga ctgcaatccc agaataaagt attggtttta accaatcaat atagtacagt
     1801 aaacatccat ttgttttgtt gaaacgttaa acaaatctga ccaaagctat tagcttatat
     1861 aaaacaggtt tgccttctat gtagctgaaa acaccacagg cccgattttg ctactgtgta
     1921 aaacatttca gcaagatttt tttattgcat ttttttttac tgaatcgttc aaacatttta
     1981 tcattttagt ttgttcattc attgcaactg gaaaaacaac acatcacaca accgcacata
     2041 tttcagcaat aagtacaata aaacactcaa ataaaaaaaa cattttaaat ctctttgtat
     2101 ttttgaccgc tgtttcgcgt aatttcacgg taaaactctg gaaatctcca ctacattcct
     2161 ctcagcggct cctctcaatg acagctgaag aagtgacgcg gctgcctgct gtgttttgat
     2221 tggtcgaatt cactggaggc ttccagaaca gtgtagagtc tgaacgggtg cgcgctctgc
     2281 tgtatttaaa gggcgaaaga gagaccgcag agaaactcaa ccgaagagaa gcgacttgac
     2341 aaagaagaaa agagcagcct gacaggactt tttccccgac gaggtgttta ttcgctctat
     2401 ttaagaatct actgtaaggt aagtctcaat atattgtact ctattggcta atcagaatta
     2461 tatagagatt atatgtactt aatgtcaaaa aatcaacttt gtatatgtaa tctttttaca
     2521 tgtggactgc ctatgttcat cttattttag gtctactaga aaattatatt tcccgttttc
     2581 acaataagga tttttaaaaa aagcaatgaa cagacgggca tttactttat gttgctgaca
     2641 ttattttata tgagcataat aaccataaat actagcaaat gtcctaaatg aatttgtgtt
     2701 aatgttgtct acaaaagaaa attagcgttt tacttgtaca actaataata acttggttat
     2761 taagagaatt tcacttgttg actagaaaaa tcctttcata atgaaacaat tgcaccataa
     2821 attgtataaa tataaaatta attctaattg tttttttttc ctgccaagtt tgtacaaaaa
     2881 agcaggctgg atggtgagcg agctgattaa ggagaacatg cacatgaagc tgtacatgga
     2941 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
     3001 gggcacccag accatgagaa tcaaggcggt cgagggcggc cctctcccct tcgccttcga
     3061 catcctggct accagcttca tgtacggcag caaaaccttc atcaaccaca cccagggcat
     3121 ccccgacttc tttaagcagt ccttccccga gggcttcaca tgggagagag tcaccacata
     3181 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ctccaggacg gctgcctcat
     3241 ctacaacgtc aagatcagag gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     3301 aacactcggc tgggaggcct ccaccgagac cctgtacccc gctgacggcg gcctggaagg
     3361 cagagccgac atggccctga agctcgtggg cgggggccac ctgatctgca acttgaagac
     3421 cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcgtct actatgtgga
     3481 cagaagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgaggt
     3541 ggctgtggcc agatactgcg acctccctag caaactgggg cacagaagat ctcgtggatc
     3601 taagctgaac cctcctgatg agagtggccc cggctgcatg agctgcaagt gtgtgctctc
     3661 ctgagaccca gctttcttgt acaaagttga tgtagatcca gacatgataa gatacattga
     3721 tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg
     3781 tgatgctatt gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa
     3841 ttgcattcat tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc
     3901 cgcggcgcca atgcattggg ccctcgagca tatgagatct gtcccgatct ttctcttctg
     3961 tgctgtcaat ttagagtagt ttttgaggta atttgggtgc attctctgta aaaacaacaa
     4021 ataaatattt atattgtatg caataaatgt aacaattaca catacactct ctgattacag
     4081 cattaaaggg ttagttcacc caaaaatgaa aagatgtcat taatgactca ccctcatgtc
     4141 gttccaagcc cgtaacacct ccgttcatct tcggaacaca gtttaagata ttttagattt
     4201 agtccgagag ctttctgtcc ctccactgaa aatgtatgta cggtatactg tccatgtcca
     4261 gaaaggtaat aaaaacatca aagtagtcca tgtgacatca gtgggttagg tagaattatt
     4321 tgaagcatcg aaaatacatt ttggtccaaa aataacaaaa cctacgactt tattcagcat
     4381 tgtagcgtca ctgcgaagtc gtgaacgcgg attgacaaca gacccggaag agaatacaat
     4441 gccgaataaa gtcgtaggtt ttgttatttt tggaccaaaa tgtattcgat gcttcaaaaa
     4501 attctaacta acccactgat gtcacatgga ctactttgat gtttttatta cctttctgga
     4561 catggacagt ataccgtaca tacattctca atggaggcac agaaagctct cggactaaat
     4621 ctaaaatatc ttaaactgtg ttctgaagat gaacggaggt cttacgggct tggaacgaca
     4681 tgagggcgag tcattaatga cattattttc atttttgggt gaactaaccc tttaataaaa
     4741 ctgacatttg cagccaattg cacactttat aatgttaaat acttgttgat tttagaaata
     4801 cagtagcatt tagtgacaat caaacattaa ttttatttct gattatattg acaaaaacac
     4861 aggactgtaa ggctgacatg acagggtgaa ttgatttcat tacattagtt ttgatggtaa
     4921 cacttttaca ataaggttcg ttagttaact acattagtta acatgaacta ataatgaact
     4981 gcacgtatac atgttaattt caacatttac taatacttta tatcttgtta acattagtta
     5041 atgcactgtg aactaacatg aacaaacaat gaacatctgt atttctatta actaacgtta
     5101 acaaagatta acagatacag taacaaatgt attgctcatg gttatttaat gttagttaat
     5161 acatgaacta atcaacctca ttgtaaagtg ttaccgtttt gacaatacaa catgtttgga
     5221 aaaaaatttc tcgcttggtt ttcttgtgtt ttgcatgaat tcaaacacaa tgagagcacg
     5281 gaatgataat gattattcaa agctgcaata ggcgccaaat ttcccgtgga cctaacgtta
     5341 ccaattacaa atgcatttta gcattttatt acgtgagcct tatattgtca atcgcatgca
     5401 ttagtgaaaa taggcctact acacatataa aaggcgcgcc atcaacgtca aaacatgcac
     5461 acccaaaaaa aaaaaaaaaa aaaaaaaaaa aaactatcag taaaacaaaa tacttaatgt
     5521 acttacctca atatgcttcc ttaggtttga tggcgaactt ttgaaggccg atatttcttt
     5581 attaagcggg agacagagga cacatttcat cttgaatgaa tctttattta caccacttaa
     5641 agaaaagaat tcgcgaagat acggccacgg gtgctcttga tcctgtggct gattttggac
     5701 tgtgctgctc gcagctgctg atgaatcaca tacttcctcc attttcttcc actgattgac
     5761 tgttataatt tccctaattt ccaggtcaag gtgctgtgca ttgtggtaat agatgtgaca
     5821 tgacgtcact tccaaaggac caatgaacat gtctgaccaa tttcatataa tgtgaaaacg
     5881 attttcatag gcagaataaa taacatttaa attaaactgg gcatcagcgc aattcaattg
     5941 gtttggtaat agcaagggaa aatagaatga agtgatctcc aaaaaataag tactttttga
     6001 ctgtaaataa aattgtaagg agtaaaaagt actttttttt ctaaaaaaat gtaattaagt
     6061 aaaagtaaaa gtattgattt ttaattgtac tcaagtaaag taaaaatccc caaaaataat
     6121 acttaagtac agtaatcaag taaaattact caagtacttt acacctctgg ttcttgaccc
     6181 cctaccttca gcaagcccag cagatccact agttctagag cggccgccac cgcggtggag
     6241 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata
     6301 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     6361 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     6421 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     6481 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     6541 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     6601 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     6661 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     6721 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     6781 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     6841 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     6901 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     6961 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     7021 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     7081 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     7141 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     7201 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     7261 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     7321 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     7381 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     7441 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7501 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7561 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     7621 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7681 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7741 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     7801 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     7861 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     7921 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     7981 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8041 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8101 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8161 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8221 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8281 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     8341 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     8401 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
//
