LOCUS       sequence-203904-        4854 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 4854)
  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 4854)
  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..4854
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..19
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1..18
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      63..72
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             69..764
                     /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    872..1006
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(896..915)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     950..969
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1119..1139)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1119..1137)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1158..1174)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1158..1174)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1171..1193)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1182..1198
                     /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(1206..1212,1213..1230,1231..1236))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1251..1272
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1389..1406)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1560..2148)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1640..1659)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2319..3110,3111..3179))
                     /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     2942..2961
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3180..3284)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3310..3765)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3447..3468)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3659..3678
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3836..4820
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 atttaggtga cactatagaa tacaagctac ttgttctttt tgcaggatct aattaggatc
       61 ccgccaccat ggtgagcgag ctgattaagg agaacatgca catgaagctg tacatggagg
      121 gcaccgtgaa caaccaccac ttcaagtgca catccgaggg cgaaggcaag ccctacgagg
      181 gcacccagac catgagaatc aaggcggtcg agggcggccc tctccccttc gccttcgaca
      241 tcctggctac cagcttcatg tacggcagca aaaccttcat caaccacacc cagggcatcc
      301 ccgacttctt taagcagtcc ttccccgagg gcttcacatg ggagagagtc accacatacg
      361 aagacggggg cgtgctgacc gctacccagg acaccagcct ccaggacggc tgcctcatct
      421 acaacgtcaa gatcagaggg gtgaacttcc catccaacgg ccctgtgatg cagaagaaaa
      481 cactcggctg ggaggcctcc accgagaccc tgtaccccgc tgacggcggc ctggaaggca
      541 gagccgacat ggccctgaag ctcgtgggcg ggggccacct gatctgcaac ttgaagacca
      601 catacagatc caagaaaccc gctaagaacc tcaagatgcc cggcgtctac tatgtggaca
      661 gaagactgga aagaatcaag gaggccgaca aagagaccta cgtcgagcag cacgaggtgg
      721 ctgtggccag atactgcgac ctccctagca aactggggca cagaagatct cgtggatcta
      781 agctgaaccc tcctgatgag agtggccccg gctgcatgag ctgcaagtgt gtgctctcct
      841 gatctagaac tatagtgagt cgtattacgt agatccagac atgataagat acattgatga
      901 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
      961 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1021 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc
     1081 ggcgccaatg cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg
     1141 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1201 ccacacaaca tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1261 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1321 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     1381 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     1441 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     1501 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     1561 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     1621 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     1681 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     1741 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     1801 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     1861 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     1921 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     1981 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     2041 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     2101 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     2161 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     2221 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     2281 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     2341 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     2401 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     2461 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     2521 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     2581 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     2641 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     2701 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     2761 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     2821 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     2881 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     2941 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     3001 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     3061 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     3121 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     3181 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     3241 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     3301 gtgccaccta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa
     3361 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
     3421 agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg
     3481 tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac
     3541 catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta
     3601 aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag
     3661 ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg
     3721 taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca
     3781 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga
     3841 ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
     3901 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
     3961 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact
     4021 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
     4081 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     4141 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     4201 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     4261 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     4321 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     4381 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     4441 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     4501 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     4561 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     4621 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     4681 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     4741 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     4801 gcaatgctcg tttagggaac cgccattctg cctggggacg tcggagcaag cttg
//
