LOCUS       sequence-451128-        2118 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MTK234 part containing lac-inducible variant of the hU6 promoter 
            driving a B to A pegRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2118)
  AUTHORS   Loveless BT, Carlson CK, Dentzel Helmy CA, Hu VJ, Ross SK, Demelo 
            MC, Murtaza A, Liang G, Ficht M, Singhai A, Pajoh-Casco MJ, Liu CC
  TITLE     Open-ended molecular recording of sequential cellular events into 
            DNA
  JOURNAL   bioRxiv 2021.11.05.467507
REFERENCE   2  (bases 1 to 2118)
  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"; journalName: "bioRxiv 
            2021.11.05.467507"
COMMENT     Sequence Label: pTBL3428 MTK234-LacO-BtoApegRNA
FEATURES             Location/Qualifiers
     source          1..2118
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             649..1308
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(715..734)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1379..1413
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    1662..1681
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     complement(1693..1712)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    1756..1775
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     complement(1862..1882)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(join(1997..2118,1..467))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2077..2096)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
       61 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
      121 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
      181 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
      241 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      301 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      361 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      421 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
      481 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
      541 tgattgatcg ggcacgtaag aggttccaac tttcaccata atgaaataag atcactaccg
      601 ggcgtatttt ttgagttatc gagattttca ggagctaagg aagctaaaat ggagaaaaaa
      661 atcactggat ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca
      721 tttcagtcag ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt
      781 ttaaagaccg taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc
      841 cgcctgatga atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata
      901 tgggatagtg ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg
      961 ctctggagtg aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg
     1021 gcgtgttacg gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc
     1081 gtttcagcca atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac
     1141 aacttcttcg cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg
     1201 atgccgctgg cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg
     1261 cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tgatatcgag
     1321 ctcgcttgga ctcctgttga tagatccagt aatgacctca gaactccatc tggatttgtt
     1381 cagaacgctc ggttgccgcc gggcgttttt tattggtctg gtctcacagc actccagtct
     1441 ttctagaaga tggaaaaagg gccatctatc attaggccat catcaccatc attgatggca
     1501 gaggaaagga agccctgctt cctccagcac cgactcggtg ccactttttc aagttgataa
     1561 cggactagcc ttatttaaac ttgctatgct gtttccagct tagctcttaa actcaatgat
     1621 agatggccca gtcggtgttt cgtcctttcc acaagatata taattgtgag cgctcacaat
     1681 tactttcaag ttacggtaag catatgatag tccattttaa aacataattt taaaactgca
     1741 aactacccaa gaaataattg tgagcgctca caattttttg tactaatatc tttgtgttta
     1801 cagtcaaatt aattctaatt atctctctaa cagccttgta tcgtatatgc aaatatgaag
     1861 gaatcatggg aaataggccc tccgtttgag accgactacg gttatccaca gaatcagggg
     1921 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1981 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     2041 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     2101 gaagctccct cgtgcgct
//
