LOCUS       sequence-376796-        6489 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for constitutive expression of LexA-CIB1-biLINuS..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6489)
  AUTHORS   Huang Z, Wu Y, Allen ME, Pan Y, Kyriakakis P, Lu S, Chang YJ, Wang 
            X, Chien S, Wang Y
  TITLE     Engineering light-controllable CAR T cells for cancer immunotherapy.
  JOURNAL   Sci Adv. 2020 Feb 19;6(8):eaay9209. doi: 10.1126/sciadv.aay9209. 
            eCollection 2020 Feb.
  PUBMED    32128416
REFERENCE   2  (bases 1 to 6489)
  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: "Sci Adv."; 
            date: "2020-02-19"; volume: "6(8):eaay9209. doi"; pages: " 
            10.1126/sciadv.aay9209. eCollection 2020 Feb"
COMMENT     Sequence Label: pCBI-LexA-CIB1-biLINuS
FEATURES             Location/Qualifiers
     source          1..6489
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        9..388
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1626..1648
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1714..1733
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     CDS             1765..2016
                     /codon_start=1
                     /gene="lexA (truncated)"
                     /product="DNA-binding domain of E. coli LexA"
                     /label=LexA BD
                     /note="binds the 16-bp palindromic sequence 
                     5'-CTGTATATATATACAG-3'"
                     /translation="MKALTARQQEVFDLIRDHISQTGMPPTRAEIAQRLGFRSPNAAEE
                     HLKALARKGVIEIVSGASRGIRLLQEEEEGLPLVGRVAA"
     primer_bind     complement(3511..3530)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3576..3631
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3630..3649)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3992..4008)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3992..4008)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4005..4027)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4016..4032
                     /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(4040..4046,4047..4064,4065..4070))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4085..4106
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4165..4360
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4211..4346
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4366..4500
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4403..4422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4457..4476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4568..4585)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4739..5327)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4819..4838)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5498..6289,6290..6358))
                     /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     6121..6140
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6359..6463)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgatcga cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
       61 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      121 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      181 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      241 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      301 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      361 ctacgtatta gtcatcgcta ttaccatggt cgaggtgagc cccacgttct gcttcactct
      421 ccccatctcc cccccctccc cacccccaat tttgtattta tttatttttt aattattttg
      481 tgcagcgatg ggggcggggg gggggggggs gcgcgccrgg gggggggggg gggggsgggg
      541 gggggggsgg ggsgrggcgg agaggtgcgg cggcagccaa tcagagcggc gcgctccgaa
      601 agtttccttt tatggcgagg cggcggcggc ggcggcccta taaaaagcga agcgcgcggc
      661 gggcgggagt cgctgcgcgc tgccttcgcc ccgtgccccg ctccgccgcc gcctcgcgcc
      721 gcccgccccg gctctgactg accgcgttac tcccacaggt gagcgggcgg gacggccctt
      781 ctcctccggg ctgtaattag cgcttggttt aatgacggct tgtttctttt ctgtggctgc
      841 gtgaaagcct tgaggggctc cgggagggcc ctttgtgcgg ggggagcggc tcggggggtg
      901 cgtgcgtgtg tgtgtgcgtg gggagcgccg cgtgcggctc cgcgctgccc ggcggctgtg
      961 agcgctgcgg gcgcggcgcg gggctttgtg cgctccgcag tgtgcgcgag gggagcgcgg
     1021 ccgggggcgg tgccccgcgg tgcggggggg gctgcgaggg gaacaaaggc tgcgtgcggg
     1081 gtgtgtgcgt gggggggtga gcagggggtg tgggcgcgtc ggtcgggctg caaccccccc
     1141 tgcacccccc tccccgagtt gctgagcacg gcccggcttc gggtgcgggg ctccgtacgg
     1201 ggcgtggcgc ggggctcgcc gtgccgggcg gggggtggcg gcaggtgggg gtgccgggcg
     1261 gggcggggcc gcctcgggcc ggggrgggct cgggggaggg gcgcggcggc ccccggagcg
     1321 ccggcggctg tcgaggcgcg gcgagccgca gccattgcct tttatggtaa tcgtgcgaga
     1381 gggcgcaggg acttcctttg tcccaaatct gtgckgrkcc saaatctgkg nngcggmgcc
     1441 gmamycyggg wggcgccgcc gcaccccctc tagcgggcgc ggggcgaagc ggtgcggcgc
     1501 cggcaggaag gaaatgggcg gggagggcct tcgtgcgtcg ccgcgccgcc gtccccttct
     1561 ccctctccag cctcggggct gtccgcgggg ggacggctgc cttcgggggg gacggggcag
     1621 ggcggggttc ggcttctggc gtgtgaccgg cggctctagc tagagcctct gctaaccatg
     1681 ttcatgcctt cttctttttc ctacagctcc tgggcaacgt gctggttatt gtgctgtctc
     1741 atcattttgg caaagaattc tgctatgaaa gcgttaacgg ccaggcaaca agaggtgttt
     1801 gatctcatcc gtgatcacat cagccagaca ggtatgccgc cgacgcgtgc ggaaatcgcg
     1861 cagcgtttgg ggttccgttc cccaaacgcg gctgaagaac atctgaaggc gctggcacgc
     1921 aaaggcgtta ttgaaattgt ttccggcgca tcacgcggga ttcgtctgtt gcaggaagag
     1981 gaagaagggt tgccgctggt aggtcgtgtg gctgccggtg aaccagctag cggcggcggc
     2041 ggcagcggcg gcggcggcag caacggcgcg attggtgggg atttgctgct taactttccc
     2101 gacatgtccg tgttggaacg tcagcgcgca catttgaagt atcttaaccc caccttcgac
     2161 tccccgttgg ccgggttctt tgcggactca tctatgatta cgggagggga aatggacagc
     2221 tacctctcaa cggccggatt gaatcttccg atgatgtatg gagaaaccac tgtagaaggc
     2281 gactcgcgac tctcgatttc gcctgaaacg acgctgggaa cagggaactt caaggcggcg
     2341 aaattcgaca cggagacaaa agattgcaac gaagcggcga agaaaatgac catgaatcgc
     2401 gatgatctgg tagaggaggg agaggaggaa aagtcgaaga ttactgaaca gaacaatggg
     2461 tctaccaaaa gtatcaaaaa gatgaagcac aaagctaaga aagaagagaa caatttcagc
     2521 aatgacagca gtaaagtcac aaaagaactg gagaaaacgg attacattca cgtgagggcg
     2581 cgacgagggc aggctacaga ttcacattca attgcggaga gagtacggag agagaaaatc
     2641 tcagaaagga tgaagttcct ccaagacctt gtgccaggtt gtgacaagat cacaggcaaa
     2701 gcaggaatgc tggatgagat catcaactac gtccaatcgt tgcaaagaca aattgagttt
     2761 ctctcgatga aactggccat cgtgaatcct agaccggatt tcgacatgga tgacatcttt
     2821 gcgaaagaag tggcatccac tcccatgacg gttgtgccct caccggagat ggtcttgtct
     2881 ggttacagcc acgaaatggt gcattcgggt tattcaagcg agatggtcaa ttcgggatac
     2941 cttcacgtca atcccatgca gcaggtgaat acttccagtg gcggcggcgg cagcggcggc
     3001 ggcggcagcg gcttggctac tacacttgaa cgtattgaga agaactttgt cattactgac
     3061 ccaagattgc cagataatcc cattatattc gcgtccgata gtttcttgca gttgacagaa
     3121 tatagccgtg aagaaatttt gggaagaaac tgcaggtttc tacaaggtcc tgaaactgat
     3181 cgcgcgacag tgagaaaaat tagagatgcc atagataacc aaacagaggt cactgttcag
     3241 ctgattaatt atacaaagag tggtaaaaag ttctggaacc tctttcactt gcagcctatg
     3301 cgagatcaga agggagatgt ccagtacttt attggggttc agttggatgg aactgagcat
     3361 gtccgagatg ctgccgagag agagggagtc atgctgatta agaaaactgc agaaaatatt
     3421 gacgaggccg caaagagact gcccgacgcc aacctggcag ccgcagccaa gaagaaaaag
     3481 ctggactgat aaagtgcatg agtcgacgga tctttttccc tctgccaaaa attatgggga
     3541 catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta ttttcattgc
     3601 aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga gggcaaatca
     3661 tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc atatgctggc
     3721 tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc cccctgctgt
     3781 ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta tattttgttt
     3841 tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac tagccagatt
     3901 tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat ggagatccct
     3961 cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     4021 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     4081 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     4141 gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag tcccgcccct
     4201 aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg
     4261 actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa
     4321 gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt gtttattgca
     4381 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     4441 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggatc
     4501 cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     4561 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     4621 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4681 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4741 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4801 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4861 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4921 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4981 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5041 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5101 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     5161 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     5221 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     5281 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     5341 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     5401 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     5461 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     5521 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     5581 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgtg
     5641 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5701 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5761 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5821 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5881 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5941 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6001 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6061 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6121 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     6181 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     6241 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     6301 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     6361 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     6421 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     6481 tgccacctg
//
