LOCUS       sequence-198328-        4108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas1, Cas2 under PLTetO-1 promoter, and TetR 
            constitutively..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4108)
  AUTHORS   Sheth RU, Yim SS, Wu FL, Wang HH
  TITLE     Multiplex recording of cellular events over time on CRISPR 
            biological tape.
  JOURNAL   Science. 2017 Dec 15;358(6369):1457-1461. doi: 
            10.1126/science.aao0958. Epub 2017 Nov 23.
  PUBMED    29170279
REFERENCE   2  (bases 1 to 4108)
  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: 
            "10.1126/science.aao0958"; journalName: "Science"; date: 
            "2017-12-15- 15"; volume: "358"; issue: "6369"; pages: "1457-1461"
COMMENT     Sequence Label: pRec Delta-LacI
FEATURES             Location/Qualifiers
     source          1..4108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(19..607)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(99..118)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(695..789)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(810..1469)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1384..1403
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1470..1572)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1638..1656
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1677..1695,1696..1701,1702..1718,1719..1724,
                     1725..1750)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1677..1695
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1702..1720
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      3010..3081
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3097..3124
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3176..3210
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             3245..3871
                     /codon_start=1
                     /gene="tetR"
                     /product="tetracycline repressor TetR"
                     /label=TetR
                     /note="TetR binds to the tetracycline operator tetO to 
                     inhibit transcription. This inhibition can be relieved by 
                     adding tetracycline or doxycycline."
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     primer_bind     complement(3321..3340)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3877..3963
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
       61 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      121 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      181 ctttctccct tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc
      241 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      301 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      361 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
      421 gttcttgaag tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc
      481 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
      541 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
      601 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
      661 acgttaaggg attttggtca tgactagtgc ttggattctc accaataaaa aacgcccggc
      721 ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact ggatctatca
      781 acaggagtcc aagcgagctc gatatcaaat tacgccccgc cctgccactc atcgcagtac
      841 tgttgtaatt cattaagcat tctgccgaca tggaagccat cacagacggc atgatgaacc
      901 tgaatcgcca gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa
      961 acgggggcga agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc
     1021 cagggattgg ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg
     1081 ttttcaccgt aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg
     1141 tggtattcac tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa
     1201 gggtgaacac tatcccatat caccagctca ccgtctttca ttgccatacg aaattccgga
     1261 tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt
     1321 ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat
     1381 tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg
     1441 gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga aaatctcgat
     1501 aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt ggaacctctt
     1561 acgtgccgat caacgtctca ttttcgccag atatcgacgt ctaagaaacc attattatca
     1621 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagtccc
     1681 tatcagtgat agagattgac atccctatca gtgatagaga tactgagcac atcagcagga
     1741 cgcactgacc gtttaataaa tatatacata tcaggacaac gcctaatgac ctggcttccc
     1801 cttaatccca ttccactcaa agatcgcgtc tccatgatct ttctgcaata tgggcagatc
     1861 gatgtaatag atggcgcgtt tgtacttatc gacaagacag ggatccgcac tcatattcct
     1921 gttggctcgg ttgcctgcat catgctggaa cctggtacac gggtttcgca tgcagctgta
     1981 cgcctggctg cgcaagttgg aacattgttg gtatgggtgg gggaagcggg cgttcgtgtt
     2041 tatgcttctg gtcagcctgg aggtgcgcgt tcagataagc tgctctatca ggcaaaactt
     2101 gctctggatg aagatttgcg tctgaaggtc gtacgtaaaa tgtttgaact tcggtttgga
     2161 gaacctgcgc ctgcccggcg ctccgtagag caactcagag gtatagaagg cagtcgcgtg
     2221 cgggcaacct acgcacttct ggcgaagcaa tacggcgtga catggaatgg acgtcgctac
     2281 gatccgaaag actgggaaaa gggcgatacg atcaaccaat gcattagcgc tgcaacttcc
     2341 tgtttatacg gcgtaactga agcggcgata cttgcagctg gttatgcacc agctattggg
     2401 tttgtgcata caggaaagcc tctttccttt gtttacgata ttgcagacat cattaaattt
     2461 gacactgttg taccgaaagc ttttgagata gcgcgtcgta accctggtga gccggaccgg
     2521 gaagtccgtt tggcgtgcag ggatattttt cgcagtagta aaacattagc caaattgatt
     2581 ccgcttatag aggacgtgct tgccgctgga gaaatacaac cgccggcccc acctgaagat
     2641 gcacagcctg ttgccattcc gcttcctgtt tcactgggag atgcaggcca tcggagtagc
     2701 tgaaatgagt atgttggtcg tggtcactga aaatgtacct ccgcgcttac gaggcagatt
     2761 agccatctgg ttgttggagg tacgtgcagg ggtatatgta ggtgatgtat ccgcaaaaat
     2821 tcgtgaaatg atctgggaac aaatagctgg actggcggaa gaaggcaatg tagtgatggc
     2881 atgggcaacg aatacggaaa cgggatttga gttccagaca tttgggttaa acaggcgtac
     2941 cccggtagat ttggatggtt taaggttggt gtctttttta cctgtttgat aataatctag
     3001 accaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt
     3061 tgtttgtcgg tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc
     3121 tgcgtttata ggtacccgtg gatcctctag agtcgacctg caggcatgca agctttttac
     3181 ggctagctca gtcctaggta tagtgctagc ccagccagag aaacactctt taacaggggg
     3241 tagtatgatg tctcgtttag ataaaagtaa agtgattaac agcgcattag agctgcttaa
     3301 tgaggtcgga atcgaaggtt taacaacccg taaactcgcc cagaagctag gtgtagagca
     3361 gcctacattg tattggcatg taaaaaataa gcgggctttg ctcgacgcct tagccattga
     3421 gatgttagat aggcaccata ctcacttttg ccctttagaa ggggaaagct ggcaagattt
     3481 tttacgtaat aacgctaaaa gttttagatg tgctttacta agtcatcgcg atggagcaaa
     3541 agtacattta ggtacacggc ctacagaaaa acagtatgaa actctcgaaa atcaattagc
     3601 ctttttatgc caacaaggtt tttcactaga gaatgcatta tatgcactca gcgcagtggg
     3661 gcattttact ttaggttgcg tattggaaga tcaagagcat caagtcgcta aagaagaaag
     3721 ggaaacacct actactgata gtatgccgcc attattacga caagctatcg aattatttga
     3781 tcaccaaggt gcagagccag ccttcttatt cggccttgaa ttgatcatat gcggattaga
     3841 aaaacaactt aaatgtgaaa gtgggtctta aggcatcaaa taaaacgaaa ggctcagtcg
     3901 aaagactggg cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca
     3961 aatccgccgc cctagaccta ggcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4021 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4081 gccagcaaaa ggccaggaac cgtaaaaa
//
