LOCUS       sequence-201414-        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"
     primer_bind     139..157
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(178..196,197..202,203..219,220..225,226..251)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    178..196
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    203..221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1511..1582
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1598..1625
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1677..1711
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1746..2372
                     /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(1822..1841)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2378..2464
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        2500..2577
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(2597..2616)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      complement(3733..4321)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3813..3832)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4409..4503)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4524..5183)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5098..5117
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 taactcaaaa aatacgaccg gtagtgatct tatttcatta tggtgaaagt tggaacctct
       61 tacgggccgg ccaacgtctc attttcgcca gatatcgacg tctaagaaac cattattatc
      121 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca cctcgagtcc
      181 ctatcagtga tagagattga catccctatc agtgatagag atactgagca catcagcagg
      241 acgcactgac cgtttaataa atatatacat atcaggacaa cgcctaatga cctggcttcc
      301 ccttaatccc attccactca aagatcgcgt ctccatgatc tttctgcaat atgggcagat
      361 cgatgtaata gatggcgcgt ttgtacttat cgacaagaca gggatccgca ctcatattcc
      421 tgttggctcg gttgcctgca tcatgctgga acctggtaca cgggtttcgc atgcagctgt
      481 acgcctggct gcgcaagttg gaacattgtt ggtatgggtg ggggaagcgg gcgttcgtgt
      541 ttatgcttct ggtcagcctg gaggtgcgcg ttcagataag ctgctctatc aggcaaaact
      601 tgctctggat gaagatttgc gtctgaaggt cgtacgtaaa atgtttgaac ttcggtttgg
      661 agaacctgcg cctgcccggc gctccgtaga gcaactcaga ggtatagaag gcagtcgcgt
      721 gcgggcaacc tacgcacttc tggcgaagca atacggcgtg acatggaatg gacgtcgcta
      781 cgatccgaaa gactgggaaa agggcgatac gatcaaccaa tgcattagcg ctgcaacttc
      841 ctgtttatac ggcgtaactg aagcggcgat acttgcagct ggttatgcac cagctattgg
      901 gtttgtgcat acaggaaagc ctctttcctt tgtttacgat attgcagaca tcattaaatt
      961 tgacactgtt gtaccgaaag cttttgagat agcgcgtcgt aaccctggtg agccggaccg
     1021 ggaagtccgt ttggcgtgca gggatatttt tcgcagtagt aaaacattag ccaaattgat
     1081 tccgcttata gaggacgtgc ttgccgctgg agaaatacaa ccgccggccc cacctgaaga
     1141 tgcacagcct gttgccattc cgcttcctgt ttcactggga gatgcaggcc atcggagtag
     1201 ctgaaatgag tatgttggtc gtggtcactg aaaatgtacc tccgcgctta cgaggcagat
     1261 tagccatctg gttgttggag gtacgtgcag gggtatatgt aggtgatgta tccgcaaaaa
     1321 ttcgtgaaat gatctgggaa caaatagctg gactggcgga agaaggcaat gtagtgatgg
     1381 catgggcaac gaatacggaa acgggatttg agttccagac atttgggtta aacaggcgta
     1441 ccccggtaga tttggatggt ttaaggttgg tgtctttttt acctgtttga taataatcta
     1501 gaccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1561 ttgtttgtcg gtgaacgctc tctactagag taacactggc tcaccttcgg gtgggccttt
     1621 ctgcgtttat aggtacccgt ggatcctcta gagtcgacct gcaggcatgc aagcttttta
     1681 cggctagctc agtcctaggt atagtgctag cccagccaga gaaacactct ttaacagggg
     1741 gtagtatgat gtctcgttta gataaaagta aagtgattaa cagcgcatta gagctgctta
     1801 atgaggtcgg aatcgaaggt ttaacaaccc gtaaactcgc ccagaagcta ggtgtagagc
     1861 agcctacatt gtattggcat gtaaaaaata agcgggcttt gctcgacgcc ttagccattg
     1921 agatgttaga taggcaccat actcactttt gccctttaga aggggaaagc tggcaagatt
     1981 ttttacgtaa taacgctaaa agttttagat gtgctttact aagtcatcgc gatggagcaa
     2041 aagtacattt aggtacacgg cctacagaaa aacagtatga aactctcgaa aatcaattag
     2101 cctttttatg ccaacaaggt ttttcactag agaatgcatt atatgcactc agcgcagtgg
     2161 ggcattttac tttaggttgc gtattggaag atcaagagca tcaagtcgct aaagaagaaa
     2221 gggaaacacc tactactgat agtatgccgc cattattacg acaagctatc gaattatttg
     2281 atcaccaagg tgcagagcca gccttcttat tcggccttga attgatcata tgcggattag
     2341 aaaaacaact taaatgtgaa agtgggtctt aaggcatcaa ataaaacgaa aggctcagtc
     2401 gaaagactgg gcctttcgtt ttatctgttg tttgtcggtg aacgctctcc tgagtaggac
     2461 aaatccgccg ccctagacga agcggcatgc atttacgttg acaccatcga atggcgcaaa
     2521 acctttcgcg gtatggcatg atagcgcccg gaagagagtc aattcagggt ggtgaatgtg
     2581 aaaccagtaa cgttatacga tgtcgcagag tatgccggtg tctcttatca gaccgtttcc
     2641 cgcgtggtga accaggccag ccacgtttct gcgaaaacgc gggaaaaagt ggaagcggcg
     2701 atggcggagc tgaattacat tcccaaccgc gcggcacaac aactggcggg caaacagtcg
     2761 aaagtggtcg ccatcattgt tacccgcctg gattcgttgt cagaaaatct cgccgttcaa
     2821 accatgctgc cagcgttcta tgaacaaggt tacgacccaa tcatgatgga aagtcagttt
     2881 tccccgcaat tagttgccga acatttgggg gtgctgaaac ggcgtaatat cgacggcgta
     2941 gtgctgttcg gttttactgg cataacagaa gaaatgttag cccactggca gtcatcgctg
     3001 gttctgctgg cgcgtgacgc aaaaggtttt gcttcggtct gttatgacga cgaaggggca
     3061 atcaagatcc tgatgcaacg gctgtatgac caggggcatc gtaatatcag ttatctcggc
     3121 gtgccgcata gtgacgtgac gaccgggaag cgacgtcacg aagcctacct ggcgttctgc
     3181 aaagcgcata aactgcatcc cgttgccgct ctgccaggcc ttgctatgaa gcaaggctat
     3241 gagaacgtag caaaagtgat tacgcctgaa actaccgcct tactgtgcgc aaccgacacg
     3301 ctggcacttg gcgcaagtaa atacctgcaa gagcaacgca tcgacacctt gcaactggcg
     3361 agcgtcggta atacgccatt aatgaaattc cttcatccgg agatcgtcac cgtagatcct
     3421 ggttacgccg aagctggacg ccaggcggcc tgccagttga tcgcacaggt aaccgggcgc
     3481 agcgaaccgc aacaaatcat catccccgcc accctgtcct gagctttcta actaaaaagg
     3541 cctcccaaat cggggggcct tttttattga taacaaaacg ctcctaggcg ttcggctgcg
     3601 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     3661 cgcatgcaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     3721 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     3781 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     3841 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     3901 cccttcggga agcgtggcgc tttctcatag ctcacgatgt aggtatctca gttcggtgta
     3961 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     4021 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     4081 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     4141 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     4201 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     4261 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     4321 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     4381 agggattttg gtcatgacta gtgcttggat tctcaccaat aaaaaacgcc cggcggcaac
     4441 cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct atcaacagga
     4501 gtccaagcga gctcgatatc aaattacgcc ccgccctgcc actcatcgca gtactgttgt
     4561 aattcattaa gcattctgcc gacatggaag ccatcacaaa cggcatgatg aacctgaatc
     4621 gccagcggca tcagcacctt gtcgccttgc gtataatatt tgcccatggt gaaaacgggg
     4681 gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac tggtgaaact cacccaggga
     4741 ttggctgaga cgaaaaacat attctcaata aaccctttag ggaaataggc caggttttca
     4801 ccataacacg ccacatcttg cgaatatatg tgtagaaact gccggaaatc gtcgtggtat
     4861 tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga aaacggtgta acaagggtga
     4921 acactatccc atatcaccag ctcaccgtct ttcattgcca tacgaaattc cggatgagca
     4981 ttcatcaggc gggcaagaat gtgaataaag gccggataaa acttgtgctt atttttcttt
     5041 acggtcttta aaaaggccgt aatatccagc tgaacggtct ggttataggt acattgagca
     5101 actgactgaa atgcctcaaa atgttcttta cgatgccatt gggatatatc aacggtggta
     5161 tatccagtga tttttttctc cattttagct tccttagctc ctgaaaatct cga
//
