LOCUS       sequence-313786-        5576 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 5576)
  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 5576)
  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
FEATURES             Location/Qualifiers
     source          1..5576
                     /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"
     polyA_signal    2606..2740
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2630..2649)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2684..2703
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(2853..2873)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2853..2871)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2892..2908)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2892..2908)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2905..2927)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2916..2932
                     /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(2940..2946,2947..2964,2965..2970))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2985..3006
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3123..3140)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3294..3882)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3374..3393)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4053..4844,4845..4913))
                     /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     4676..4695
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4914..5018)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5044..5499)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5181..5202)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5393..5412
                     /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 ctgtagtcta
     2581 gaactatagt gagtcgtatt acgtagatcc agacatgata agatacattg atgagtttgg
     2641 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     2701 tgctttattt gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca
     2761 ttttatgttt caggttcagg gggaggtgtg ggaggttttt taattcgcgg ccgcggcgcc
     2821 aatgcattgg gcccggtacc cagcttttgt tccctttagt gagggttaat tgcgcgcttg
     2881 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     2941 aacatacgag ccgggagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3001 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3061 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3121 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     3181 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     3241 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     3301 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     3361 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     3421 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     3481 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     3541 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     3601 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     3661 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     3721 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     3781 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     3841 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     3901 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     3961 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4021 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4081 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4141 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     4201 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     4261 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     4321 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     4381 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     4441 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     4501 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     4561 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     4621 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     4681 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     4741 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     4801 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     4861 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     4921 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     4981 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     5041 cctaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     5101 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     5161 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     5221 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     5281 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     5341 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     5401 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     5461 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca ttcaggctgc
     5521 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag tcgacc
//
