LOCUS       sequence-451127-        2114 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MTK234 part containing tet-inducible variant of the hU6 promoter 
            driving an A to B pegRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2114)
  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 2114)
  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: pTBL3331 MTK234-TetO-AtoBpegRNA
FEATURES             Location/Qualifiers
     source          1..2114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      474..508
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     protein_bind    726..744
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    754..772
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(783..802)
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     primer_bind     complement(953..973)
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     rep_origin      complement(1088..1676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1168..1187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1858..2114,1..403)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1924..1943)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
ORIGIN
        1 tagtgttcac ccttgttaca ccgttttcca tgagcaaact gaaacgtttt catcgctctg
       61 gagtgaatac cacgacgatt tccggcagtt tctacacata tattcgcaag atgtggcgtg
      121 ttacggtgaa aacctggcct atttccctaa agggtttatt gagaatatgt ttttcgtttc
      181 agccaatccc tgggtgagtt tcaccagttt tgatttaaac gtggccaata tggacaactt
      241 cttcgccccc gttttcacca tgggcaaata ttatacgcaa ggcgacaagg tgctgatgcc
      301 gctggcgatt caggttcatc atgccgtttg tgatggcttc catgtcggca gaatgcttaa
      361 tgaattacaa cagtactgcg atgagtggca gggcggggcg taatttgata tcgagctcgc
      421 ttggactcct gttgatagat ccagtaatga cctcagaact ccatctggat ttgttcagaa
      481 cgctcggttg ccgccgggcg ttttttattg gtctggtctc acagcactcc agtctttcta
      541 gaagatggaa aaatcaccat cataccactg ggccatctat cattgatggc agaggaaagg
      601 aagccctgct tcctccagca ccgactcggt gccacttttt caagttgata acggactagc
      661 cttatttaaa cttgctatgc tgtttccagc ttagctctta aactcaatga tggtgatgat
      721 ggcggtctct atcactgata gggagatata taatctctat cactgatagg gagtttcaag
      781 ttacggtaag catatgatag tccattttaa aacataattt taaaactgca aactacccaa
      841 gaaattatta ctttctacgt cacgtatttt gtactaatat ctttgtgttt acagtcaaat
      901 taattctaat tatctctcta acagccttgt atcgtatatg caaatatgaa ggaatcatgg
      961 gaaataggcc ctccgtttga gaccgactac ggttatccac agaatcaggg gataacgcag
     1021 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1081 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     1141 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     1201 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     1261 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     1321 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     1381 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     1441 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     1501 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     1561 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     1621 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     1681 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     1741 ttttggtcat gattgatcgg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
     1801 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
     1861 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
     1921 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
     1981 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
     2041 attcttgccc gcctgatgaa tgctcatccg gaatttcgta tggcaatgaa agacggtgag
     2101 ctggtgatat ggga
//
