LOCUS       sequence-201412-        5315 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Cas1, Cas2 under PLTetO-1 promoter, and TetR and LacI-GalS
            chimera constitutively..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5315)
  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 5315)
  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-GalS
FEATURES             Location/Qualifiers
     source          1..5315
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     156..174
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(195..213,214..219,220..236,237..242,243..268)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    195..213
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    220..238
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     terminator      1528..1599
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1615..1642
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        1694..1728
                     /label=BBa_J23106
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.47"
     CDS             1763..2389
                     /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(1839..1858)
                     /label=Tet-R
                     /note="Tetracycline resistance gene, reverse primer"
     terminator      2395..2481
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        2517..2594
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     complement(2614..2633)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     rep_origin      complement(3852..4440)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3932..3951)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4528..4622)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(4643..5302)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     5217..5236
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 gctcctgaaa atctcgataa ctcaaaaaat acgaccggta gtgatcttat ttcattatgg
       61 tgaaagttgg aacctcttac gggccggcca acgtctcatt ttcgccagat atcgacgtct
      121 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
      181 gtcttcacct cgagtcccta tcagtgatag agattgacat ccctatcagt gatagagata
      241 ctgagcacat cagcaggacg cactgaccgt ttaataaata tatacatatc aggacaacgc
      301 ctaatgacct ggcttcccct taatcccatt ccactcaaag atcgcgtctc catgatcttt
      361 ctgcaatatg ggcagatcga tgtaatagat ggcgcgtttg tacttatcga caagacaggg
      421 atccgcactc atattcctgt tggctcggtt gcctgcatca tgctggaacc tggtacacgg
      481 gtttcgcatg cagctgtacg cctggctgcg caagttggaa cattgttggt atgggtgggg
      541 gaagcgggcg ttcgtgttta tgcttctggt cagcctggag gtgcgcgttc agataagctg
      601 ctctatcagg caaaacttgc tctggatgaa gatttgcgtc tgaaggtcgt acgtaaaatg
      661 tttgaacttc ggtttggaga acctgcgcct gcccggcgct ccgtagagca actcagaggt
      721 atagaaggca gtcgcgtgcg ggcaacctac gcacttctgg cgaagcaata cggcgtgaca
      781 tggaatggac gtcgctacga tccgaaagac tgggaaaagg gcgatacgat caaccaatgc
      841 attagcgctg caacttcctg tttatacggc gtaactgaag cggcgatact tgcagctggt
      901 tatgcaccag ctattgggtt tgtgcataca ggaaagcctc tttcctttgt ttacgatatt
      961 gcagacatca ttaaatttga cactgttgta ccgaaagctt ttgagatagc gcgtcgtaac
     1021 cctggtgagc cggaccggga agtccgtttg gcgtgcaggg atatttttcg cagtagtaaa
     1081 acattagcca aattgattcc gcttatagag gacgtgcttg ccgctggaga aatacaaccg
     1141 ccggccccac ctgaagatgc acagcctgtt gccattccgc ttcctgtttc actgggagat
     1201 gcaggccatc ggagtagctg aaatgagtat gttggtcgtg gtcactgaaa atgtacctcc
     1261 gcgcttacga ggcagattag ccatctggtt gttggaggta cgtgcagggg tatatgtagg
     1321 tgatgtatcc gcaaaaattc gtgaaatgat ctgggaacaa atagctggac tggcggaaga
     1381 aggcaatgta gtgatggcat gggcaacgaa tacggaaacg ggatttgagt tccagacatt
     1441 tgggttaaac aggcgtaccc cggtagattt ggatggttta aggttggtgt cttttttacc
     1501 tgtttgataa taatctagac caggcatcaa ataaaacgaa aggctcagtc gaaagactgg
     1561 gcctttcgtt ttatctgttg tttgtcggtg aacgctctct actagagtaa cactggctca
     1621 ccttcgggtg ggcctttctg cgtttatagg tacccgtgga tcctctagag tcgacctgca
     1681 ggcatgcaag ctttttacgg ctagctcagt cctaggtata gtgctagccc agccagagaa
     1741 acactcttta acagggggta gtatgatgtc tcgtttagat aaaagtaaag tgattaacag
     1801 cgcattagag ctgcttaatg aggtcggaat cgaaggttta acaacccgta aactcgccca
     1861 gaagctaggt gtagagcagc ctacattgta ttggcatgta aaaaataagc gggctttgct
     1921 cgacgcctta gccattgaga tgttagatag gcaccatact cacttttgcc ctttagaagg
     1981 ggaaagctgg caagattttt tacgtaataa cgctaaaagt tttagatgtg ctttactaag
     2041 tcatcgcgat ggagcaaaag tacatttagg tacacggcct acagaaaaac agtatgaaac
     2101 tctcgaaaat caattagcct ttttatgcca acaaggtttt tcactagaga atgcattata
     2161 tgcactcagc gcagtggggc attttacttt aggttgcgta ttggaagatc aagagcatca
     2221 agtcgctaaa gaagaaaggg aaacacctac tactgatagt atgccgccat tattacgaca
     2281 agctatcgaa ttatttgatc accaaggtgc agagccagcc ttcttattcg gccttgaatt
     2341 gatcatatgc ggattagaaa aacaacttaa atgtgaaagt gggtcttaag gcatcaaata
     2401 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     2461 gctctcctga gtaggacaaa tccgccgccc tagacgaagc ggcatgcatt tacgttgaca
     2521 ccatcgaatg gcgcaaaacc tttcgcggta tggcatgata gcgcccggaa gagagtcaat
     2581 tcagggtggt gaatgtgaaa ccagtaacgt tatacgatgt cgcagagtat gccggtgtct
     2641 cttatcagac cgtttcccgc gtggtgaacc aggccagcca cgtttctgcg aaaacgcggg
     2701 aaaaagtgga agcggcgatg gcggagctga attacattcc caaccgcgtg gcagcgcaac
     2761 tggcgggcaa acagtcggac accattggcg tggtggtgat ggacgtttct gatgcttttt
     2821 tcggcgcgct ggtaaaagcg gtggatctgg tcgcccagca gcatcagaaa tacgtgctaa
     2881 tcggcaatag ctatcatgaa gcggaaaaag agcgtcacgc cattgaggtg ttaattcgcc
     2941 agcgttgtaa tgcgttgatt gttcactcaa aagcattgag tgacgatgaa ctggcgcaat
     3001 ttatggataa cattcccggt atggtgttaa tcaaccgcgt tgtgccgggg tacgcccatc
     3061 gttgcgtttg cctggataat ctcagcggtg cccgaatggc gacgcgcatg ttgctgaata
     3121 acggtcatca acgtattggt tatctttctt ccagccacgg cattgaagat gacgccatgc
     3181 gtaaagcagg ctggatgagt gcgttgaaag agcaggatat tattccgcca gaaagctgga
     3241 ttggcactgg tacgccggac atgccgggcg gtgaggcggc gatggttaaa ctgctggggc
     3301 gcaatctaca acttaccgct gtatttgctt ataacgacaa tatggccgct ggcgcactga
     3361 cagcattaaa agataatggc attgcgattc cgttacatct ctcaatcatc ggtttcgatg
     3421 atattcccat cgcccgttac accgacccgc aattaacgac cgtgcgttat cccattgctt
     3481 caatggcgaa attagccacc gaactggcct tgcagggggc agcaggcaat attgatcctc
     3541 gtgccagcca ctggtttatg ccgacgttag tgcgtcgcca ttctgtcgca acacgccaga
     3601 atgcggcggc gatcactaac tcaacaaatc aggcgatgta agctttctaa ctaaaaaggc
     3661 ctcccaaatc ggggggcctt ttttattgat aacaaaacgc tcctaggcgt tcggctgcgg
     3721 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3781 gcatgcaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     3841 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3901 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3961 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4021 ccttcgggaa gcgtggcgct ttctcatagc tcacgatgta ggtatctcag ttcggtgtag
     4081 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4141 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4201 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4261 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     4321 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4381 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4441 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4501 gggattttgg tcatgactag tgcttggatt ctcaccaata aaaaacgccc ggcggcaacc
     4561 gagcgttctg aacaaatcca gatggagttc tgaggtcatt actggatcta tcaacaggag
     4621 tccaagcgag ctcgatatca aattacgccc cgccctgcca ctcatcgcag tactgttgta
     4681 attcattaag cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg
     4741 ccagcggcat cagcaccttg tcgccttgcg tataatattt gcccatggtg aaaacggggg
     4801 cgaagaagtt gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat
     4861 tggctgagac gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac
     4921 cataacacgc cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt
     4981 cactccagag cgatgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa
     5041 cactatccca tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat
     5101 tcatcaggcg ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta
     5161 cggtctttaa aaaggccgta atatccagct gaacggtctg gttataggta cattgagcaa
     5221 ctgactgaaa tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat
     5281 atccagtgat ttttttctcc attttagctt cctta
//
