LOCUS       sequence-313784-        6294 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For use in light-induced protein inactivation through the 
            interaction with CIB1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6294)
  AUTHORS   Rathbun LI, Colicino EG, Manikas J, O'Connell J, Krishnan N, Reilly 
            NS, Coyne S, Erdemci-Tandogan G, Garrastegui A, Freshour J, Santra 
            P, Manning ML, Amack JD, Hehnly H
  TITLE     Cytokinetic bridge triggers de novo lumen formation in vivo.
  JOURNAL   Nat Commun. 2020 Mar 9;11(1):1269. doi: 10.1038/s41467-020-15002-8.
  PUBMED    32152267
REFERENCE   2  (bases 1 to 6294)
  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"; journalName: "Nat 
            Commun."; date: "2020-03-9"; volume: "11(1):1269. doi"; pages: " 
            10.1038/s41467-020-15002-8"
COMMENT     Sequence Label: pCS2-CRY2-mCherry
FEATURES             Location/Qualifiers
     source          1..6294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..978
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1013..1031
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1013..1030
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      2574..2583
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2580..3287
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2725..2743)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2993..3012
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3324..3458
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3348..3367)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3402..3421
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3571..3591)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3571..3589)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3610..3626)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3610..3626)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3623..3645)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3634..3650
                     /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(3658..3664,3665..3682,3683..3688))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3703..3724
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3841..3858)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4012..4600)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4092..4111)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4771..5562,5563..5631))
                     /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     5394..5413
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5632..5736)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5762..6217)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5899..5920)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6111..6130
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gccaattcaa tatggcgtat atggactcat gccaattcaa tatggtggat ctggacctgt
       61 gccaattcaa tatggcgtat atggactcgt gccaattcaa tatggtggat ctggacccca
      121 gccaattcaa tatggcggac ttggcaccat gccaattcaa tatggcggac ctggcactgt
      181 gccaactggg gaggggtcta cttggcacgg tgccaagttt gaggaggggt cttggccctg
      241 tgccaagtcc gccatattga attggcatgg tgccaataat ggcggccata ttggctatat
      301 gccaggatca atatataggc aatatccaat atggccctat gccaatatgg ctattggcca
      361 ggttcaatac tatgtattgg ccctatgcca tatagtattc catatatggg ttttcctatt
      421 gacgtagata gcccctccca atgggcggtc ccatatacca tatatggggc ttcctaatac
      481 cgcccatagc cactccccca ttgacgtcaa tggtctctat atatggtctt tcctattgac
      541 gtcatatggg cggtcctatt gacgtatatg gcgcctcccc cattgacgtc aattacggta
      601 aatggcccgc ctggctcaat gcccattgac gtcaatagga ccacccacca ttgacgtcaa
      661 tgggatggct cattgcccat tcatatccgt tctcacgccc cctattgacg tcaatgacgg
      721 taaatggccc acttggcagt acatcaatat ctattaatag taacttggca agtacattac
      781 tattggaagt acgccagggt acattggcag tactcccatt gacgtcaatg gcggtaaatg
      841 gcccgcgatg gctgccaagt acatccccat tgacgtcaat ggggaggggc aatgacgcaa
      901 atgggcgttc cattgacgta aatgggcggt aggcgtgcct aatgggaggt ctatataagc
      961 aatgctcgtt tagggaaccg ccattctgcc tggggacgtc ggagcaagct tgatttaggt
     1021 gacactatag aatacaagct acttgttctt tttgcagcat gaagatggac aaaaagacca
     1081 tcgtctggtt tcggagagat ttgagaatag aagataatcc cgcgctcgcc gccgcggccc
     1141 acgagggttc cgtcttcccc gttttcattt ggtgtcctga agaagaaggc cagttttatc
     1201 ccggaagggc ctctaggtgg tggatgaagc aaagtctggc ccatcttagc cagtcactga
     1261 aagcactggg cagtgatctt accctgatca agacacacaa taccatctct gccatcctcg
     1321 actgcatcag ggtgaccggc gcaacgaaag tcgtgtttaa ccacctgtac gatccagtta
     1381 gtctggtgcg cgaccacact gtgaaggaga agctggtgga acgggggatc agtgtgcaga
     1441 gctacaacgg ggaccttctg tacgagccat gggagatcta ttgcgagaaa gggaaaccgt
     1501 tcacctcctt caacagttac tggaagaaat gtttggatat gtcaatagag tccgttatgt
     1561 tgccccctcc ctggagactg atgccgatta ctgctgctgc agaggccatc tgggcctgct
     1621 ccatcgagga actcggtctg gaaaatgaag cagaaaagcc aagcaatgca cttctcacta
     1681 gagcctggag ccccggctgg tctaatgccg acaagctgct taacgagttc atcgaaaaac
     1741 aactgattga ctacgcgaag aactccaaga aagtggtagg taactcaact agcttgctct
     1801 ctccatatct ccattttggc gagatttctg tccgccatgt atttcagtgc gctcggatga
     1861 aacagattat ctgggctcgc gataaaaaca gcgaaggcga agaaagcgcc gatctgttcc
     1921 tgcgagggat cggacttcgg gaatactccc ggtatatatg tttcaacttt ccattcacac
     1981 acgagcagag tctgttgtcc cacctcaggt tcttcccctg ggacgccgat gtcgacaaat
     2041 tcaaggcatg gagacaggga aggacagggt acccactcgt ggatgctggc atgagagagc
     2101 tctgggctac aggctggatg cacaaccgca tccgggtaat cgtgtcctca tttgctgtca
     2161 agtttctgct cctgccttgg aaatggggaa tgaagtactt ttgggatacc cttctcgacg
     2221 ccgacttgga gtgtgacatt ctgggatggc aatatattag cgggtcaatt cctgacggcc
     2281 atgagttgga caggttggac aatccggcct tgcagggagc taagtatgat cccgaaggag
     2341 agtatattcg acagtggctc cccgagctgg cccgacttcc tacggagtgg attcaccatc
     2401 cttgggacgc accactgaca gtgctcaagg caagcggggt ggagctgggc accaattacg
     2461 ctaagcctat agttgatata gatacagcac gcgagctgct ggctaaagcg atctctcgca
     2521 ctcgggaggc gcagattatg atcggtgctg ccggcagcgg cagcccaccg gtcgccacca
     2581 tggtgagcaa gggcgaggag gataacatgg ccatcatcaa ggagttcatg cgcttcaagg
     2641 tgcacatgga gggctccgtg aacggccacg agttcgagat cgagggcgag ggcgagggcc
     2701 gcccctacga gggcacccag accgccaagc tgaaggtgac caagggtggc cccctgccct
     2761 tcgcctggga catcctgtcc cctcagttca tgtacggctc caaggcctac gtgaagcacc
     2821 ccgccgacat ccccgactac ttgaagctgt ccttccccga gggcttcaag tgggagcgcg
     2881 tgatgaactt cgaggacggc ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg
     2941 gcgagttcat ctacaaggtg aagctgcgcg gcaccaactt cccctccgac ggccccgtaa
     3001 tgcagaagaa gaccatgggc tgggaggcct cctccgagcg gatgtacccc gaggacggcg
     3061 ccctgaaggg cgagatcaag cagaggctga agctgaagga cggcggccac tacgacgctg
     3121 aggtcaagac cacctacaag gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca
     3181 acatcaagtt ggacatcacc tcccacaacg aggactacac catcgtggaa cagtacgaac
     3241 gcgccgaggg ccgccactcc accggcggca tggacgagct gtacaagtct gcagtctaga
     3301 actatagtga gtcgtattac gtagatccag acatgataag atacattgat gagtttggac
     3361 aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt gatgctattg
     3421 ctttatttgt aaccattata agctgcaata aacaagttaa caacaacaat tgcattcatt
     3481 ttatgtttca ggttcagggg gaggtgtggg aggtttttta attcgcggcc gcggcgccaa
     3541 tgcattgggc ccggtaccca gcttttgttc cctttagtga gggttaattg cgcgcttggc
     3601 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     3661 catacgagcc gggagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     3721 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     3781 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     3841 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     3901 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     3961 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     4021 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     4081 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     4141 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     4201 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     4261 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     4321 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     4381 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     4441 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     4501 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     4561 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     4621 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     4681 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     4741 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     4801 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     4861 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     4921 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     4981 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5041 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5101 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5161 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5221 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5281 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5341 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5401 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5461 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5521 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     5581 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     5641 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     5701 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     5761 taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca
     5821 ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag
     5881 atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc
     5941 aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga accatcaccc
     6001 taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc taaagggagc
     6061 ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa
     6121 gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc
     6181 acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt caggctgcgc
     6241 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagtc gacc
//
