LOCUS       sequence-201432-        4109 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 4109)
  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 4109)
  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..4109
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(13..601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(93..112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(689..783)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(804..1463)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1378..1397
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     1632..1650
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1671..1689,1690..1695,1696..1712,1713..1718,
                     1719..1744)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1671..1689
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1696..1714
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      3004..3075
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3091..3118
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        3170..3204
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             3239..3865
                     /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(3315..3334)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      3871..3957
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
       61 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
      121 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
      181 cccttcggga agcgtggcgc tttctcatag ctcacgatgt aggtatctca gttcggtgta
      241 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
      301 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
      361 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
      421 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
      481 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
      541 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
      601 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
      661 agggattttg gtcatgacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
      721 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
      781 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
      841 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
      901 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
      961 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     1021 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     1081 ccataacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     1141 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     1201 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     1261 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     1321 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     1381 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     1441 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cgataactca
     1501 aaaaatacga ccggtagtga tcttatttca ttatggtgaa agttggaacc tcttacgggc
     1561 cggccaacgt ctcattttcg ccagatatcg acgtctaaga aaccattatt atcatgacat
     1621 taacctataa aaataggcgt atcacgaggc cctttcgtct tcacctcgag tccctatcag
     1681 tgatagagat tgacatccct atcagtgata gagatactga gcacatcagc aggacgcact
     1741 gaccgtttaa taaatatata catatcagga caacgcctaa tgacctggct tccccttaat
     1801 cccattccac tcaaagatcg cgtctccatg atctttctgc aatatgggca gatcgatgta
     1861 atagatggcg cgtttgtact tatcgacaag acagggatcc gcactcatat tcctgttggc
     1921 tcggttgcct gcatcatgct ggaacctggt acacgggttt cgcatgcagc tgtacgcctg
     1981 gctgcgcaag ttggaacatt gttggtatgg gtgggggaag cgggcgttcg tgtttatgct
     2041 tctggtcagc ctggaggtgc gcgttcagat aagctgctct atcaggcaaa acttgctctg
     2101 gatgaagatt tgcgtctgaa ggtcgtacgt aaaatgtttg aacttcggtt tggagaacct
     2161 gcgcctgccc ggcgctccgt agagcaactc agaggtatag aaggcagtcg cgtgcgggca
     2221 acctacgcac ttctggcgaa gcaatacggc gtgacatgga atggacgtcg ctacgatccg
     2281 aaagactggg aaaagggcga tacgatcaac caatgcatta gcgctgcaac ttcctgttta
     2341 tacggcgtaa ctgaagcggc gatacttgca gctggttatg caccagctat tgggtttgtg
     2401 catacaggaa agcctctttc ctttgtttac gatattgcag acatcattaa atttgacact
     2461 gttgtaccga aagcttttga gatagcgcgt cgtaaccctg gtgagccgga ccgggaagtc
     2521 cgtttggcgt gcagggatat ttttcgcagt agtaaaacat tagccaaatt gattccgctt
     2581 atagaggacg tgcttgccgc tggagaaata caaccgccgg ccccacctga agatgcacag
     2641 cctgttgcca ttccgcttcc tgtttcactg ggagatgcag gccatcggag tagctgaaat
     2701 gagtatgttg gtcgtggtca ctgaaaatgt acctccgcgc ttacgaggca gattagccat
     2761 ctggttgttg gaggtacgtg caggggtata tgtaggtgat gtatccgcaa aaattcgtga
     2821 aatgatctgg gaacaaatag ctggactggc ggaagaaggc aatgtagtga tggcatgggc
     2881 aacgaatacg gaaacgggat ttgagttcca gacatttggg ttaaacaggc gtaccccggt
     2941 agatttggat ggtttaaggt tggtgtcttt tttacctgtt tgataataat ctagaccagg
     3001 catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat ctgttgtttg
     3061 tcggtgaacg ctctctacta gagtaacact ggctcacctt cgggtgggcc tttctgcgtt
     3121 tataggtacc cgtggatcct ctagagtcga cctgcaggca tgcaagcttt ttacggctag
     3181 ctcagtccta ggtatagtgc tagcccagcc agagaaacac tctttaacag ggggtagtat
     3241 gatgtctcgt ttagataaaa gtaaagtgat taacagcgca ttagagctgc ttaatgaggt
     3301 cggaatcgaa ggtttaacaa cccgtaaact cgcccagaag ctaggtgtag agcagcctac
     3361 attgtattgg catgtaaaaa ataagcgggc tttgctcgac gccttagcca ttgagatgtt
     3421 agataggcac catactcact tttgcccttt agaaggggaa agctggcaag attttttacg
     3481 taataacgct aaaagtttta gatgtgcttt actaagtcat cgcgatggag caaaagtaca
     3541 tttaggtaca cggcctacag aaaaacagta tgaaactctc gaaaatcaat tagccttttt
     3601 atgccaacaa ggtttttcac tagagaatgc attatatgca ctcagcgcag tggggcattt
     3661 tactttaggt tgcgtattgg aagatcaaga gcatcaagtc gctaaagaag aaagggaaac
     3721 acctactact gatagtatgc cgccattatt acgacaagct atcgaattat ttgatcacca
     3781 aggtgcagag ccagccttct tattcggcct tgaattgatc atatgcggat tagaaaaaca
     3841 acttaaatgt gaaagtgggt cttaaggcat caaataaaac gaaaggctca gtcgaaagac
     3901 tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg
     3961 ccgccctaga cctagggcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     4021 atacggttat ccacagaatc aggggataac gcatgcaaga acatgtgagc aaaaggccag
     4081 caaaaggcca ggaaccgtaa aaaggccgc
//
