LOCUS       sequence-198327-        5213 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas1, Cas2 under PLTetO-1 promoter, and TetR and LacI-TreR
            chimera constitutively..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5213)
  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 5213)
  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-TreR
FEATURES             Location/Qualifiers
     source          1..5213
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..103)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     169..187
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(208..226,227..232,233..249,250..255,256..281)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    208..226
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    233..251
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1541..1612
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1628..1655
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1707..1741
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1776..2402
                     /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(1852..1871)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2408..2494
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        2530..2607
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     complement(2627..2646)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      complement(3763..4351)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3843..3862)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4439..4533)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4554..5213)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5128..5147
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
       61 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attttcgcca
      121 gatatcgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
      181 acgaggccct ttcgtcttca cctcgagtcc ctatcagtga tagagattga catccctatc
      241 agtgatagag atactgagca catcagcagg acgcactgac cgtttaataa atatatacat
      301 atcaggacaa cgcctaatga cctggcttcc ccttaatccc attccactca aagatcgcgt
      361 ctccatgatc tttctgcaat atgggcagat cgatgtaata gatggcgcgt ttgtacttat
      421 cgacaagaca gggatccgca ctcatattcc tgttggctcg gttgcctgca tcatgctgga
      481 acctggtaca cgggtttcgc atgcagctgt acgcctggct gcgcaagttg gaacattgtt
      541 ggtatgggtg ggggaagcgg gcgttcgtgt ttatgcttct ggtcagcctg gaggtgcgcg
      601 ttcagataag ctgctctatc aggcaaaact tgctctggat gaagatttgc gtctgaaggt
      661 cgtacgtaaa atgtttgaac ttcggtttgg agaacctgcg cctgcccggc gctccgtaga
      721 gcaactcaga ggtatagaag gcagtcgcgt gcgggcaacc tacgcacttc tggcgaagca
      781 atacggcgtg acatggaatg gacgtcgcta cgatccgaaa gactgggaaa agggcgatac
      841 gatcaaccaa tgcattagcg ctgcaacttc ctgtttatac ggcgtaactg aagcggcgat
      901 acttgcagct ggttatgcac cagctattgg gtttgtgcat acaggaaagc ctctttcctt
      961 tgtttacgat attgcagaca tcattaaatt tgacactgtt gtaccgaaag cttttgagat
     1021 agcgcgtcgt aaccctggtg agccggaccg ggaagtccgt ttggcgtgca gggatatttt
     1081 tcgcagtagt aaaacattag ccaaattgat tccgcttata gaggacgtgc ttgccgctgg
     1141 agaaatacaa ccgccggccc cacctgaaga tgcacagcct gttgccattc cgcttcctgt
     1201 ttcactggga gatgcaggcc atcggagtag ctgaaatgag tatgttggtc gtggtcactg
     1261 aaaatgtacc tccgcgctta cgaggcagat tagccatctg gttgttggag gtacgtgcag
     1321 gggtatatgt aggtgatgta tccgcaaaaa ttcgtgaaat gatctgggaa caaatagctg
     1381 gactggcgga agaaggcaat gtagtgatgg catgggcaac gaatacggaa acgggatttg
     1441 agttccagac atttgggtta aacaggcgta ccccggtaga tttggatggt ttaaggttgg
     1501 tgtctttttt acctgtttga taataatcta gaccaggcat caaataaaac gaaaggctca
     1561 gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tctactagag
     1621 tcacactggc tcaccttcgg gtgggccttt ctgcgtttat aggtacccgt ggatcctcta
     1681 gagtcgacct gcaggcatgc aagcttttta cggctagctc agtcctaggt atagtgctag
     1741 cccagccaga gaaacactct ttaacagggg gtagtatgat gtctcgttta gataaaagta
     1801 aagtgattaa cagcgcatta gagctgctta atgaggtcgg aatcgaaggt ttaacaaccc
     1861 gtaaactcgc ccagaagcta ggtgtagagc agcctacatt gtattggcat gtaaaaaata
     1921 agcgggcttt gctcgacgcc ttagccattg agatgttaga taggcaccat actcactttt
     1981 gccctttaga aggggaaagc tggcaagatt ttttacgtaa taacgctaaa agttttagat
     2041 gtgctttact aagtcatcgc gatggagcaa aagtacattt aggtacacgg cctacagaaa
     2101 aacagtatga aactctcgaa aatcaattag cctttttatg ccaacaaggt ttttcactag
     2161 agaatgcatt atatgcactc agcgcagtgg ggcattttac tttaggttgc gtattggaag
     2221 atcaagagca tcaagtcgct aaagaagaaa gggaaacacc tactactgat agtatgccgc
     2281 cattattacg acaagctatc gaattatttg atcaccaagg tgcagagcca gccttcttat
     2341 tcggccttga attgatcata tgcggattag aaaaacaact taaatgtgaa agtgggtctt
     2401 aaggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt ttatctgttg
     2461 tttgtcggtg aacgctctcc tgagtaggac aaatccgccg ccctagacga agcggcatgc
     2521 atttacgttg acaccatcga atggtgcaaa acctttcgcg gtatggcatg atagcgcccg
     2581 gaagagagtc aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag
     2641 tatgccggtg tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct
     2701 gcgaaaacgc gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc
     2761 gcggcacaac aactggcggg caaacagtcg aaagtggtcg ccatcattgt tacccgcctg
     2821 gattcgttgt cagaaaatct cgccgttcaa accatgctgc cagcgttcta tgaacaaggt
     2881 tacgacccaa tcatgatgga aagtcagttt tccccgcaat tagttgccga acatttgggg
     2941 gtgctgaaac ggcgtaatat cgacggcgta gtgctgttcg gttttactgg cataacagaa
     3001 gaaatgttag cccactggca gtcatcgctg gttctgctgg cgcgtgacgc aaaaggtttt
     3061 gcttcggtct gttatgacga cgaaggggca atcaagatcc tgatgcaacg gctgtatgac
     3121 caggggcatc gtaatatcag ttatctcggc gtgccgcata gtgacgtgac gaccgggaag
     3181 cgacgtcacg aagcctacct ggcgttctgc aaagcgcata aactgcatcc cgttgccgct
     3241 ctgccaggcc ttgctatgaa gcaaggctat gagaacgtag caaaagtgat tacgcctgaa
     3301 actaccgcct tactgtgcgc aaccgacacg ctggcacttg gcgcaagtaa atacctgcaa
     3361 gagcaacgca tcgacacctt gcaactggcg agcgtcggta atacgccatt aatgaaattc
     3421 cttcatccgg agatcgtcac cgtagatcct ggttacgccg aagctggacg ccaggcggcc
     3481 tgccagttga tcgcacaggt aaccgggcgc agcgaaccgc aacaaatcat catccccgcc
     3541 accctgtcct gagctttcta actaaaaagg cctcccaaat cggggggcct tttttattga
     3601 taacaaaacg ctcctaggcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     3661 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     3721 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3781 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3841 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3901 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     3961 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4021 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4081 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4141 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     4201 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4261 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4321 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4381 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgacta gtgcttggat
     4441 tctcaccaat aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt
     4501 ctgaggtcat tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc
     4561 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     4621 ccatcacaga cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     4681 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     4741 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     4801 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     4861 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     4921 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     4981 ttcattgcca tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     5041 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     5101 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     5161 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cat
//
