LOCUS       sequence-401268-        2687 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 part.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2687)
  AUTHORS   Gilbert C, Brumwell SL, Crits-Christoph A, Kang SC, Martin-Moldes Z,
            Alsharif W, Esmurria A, Nguyen M-A, Lee HH, Ostrov N
  TITLE     A scalable framework for high-throughput identification of 
            functional origins of replication in non-model bacteria
  JOURNAL   bioRxiv 2023.05.19.541510
REFERENCE   2  (bases 1 to 2687)
  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 
            2023.05.19.541510"
COMMENT     Sequence Label: pGL0_15 [CEN6/ARSH4 ORI]
FEATURES             Location/Qualifiers
     source          1..2687
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(420..514)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(535..1194)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1109..1128
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1195..1297)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1384..1406
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1398..1415
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1399..1415
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1552..2055
                     /label=CEN/ARS
                     /note="S. cerevisiae CEN6 centromere fused to an 
                     autonomously replicating sequence"
     primer_bind     complement(2198..2214)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2198..2214)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2211..2233)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(2386..2687,1..287))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2466..2485)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
       61 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
      121 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
      181 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
      241 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
      301 tcttttctac ggggtctgac gctctattca acaaagccgc cgtcccgtca agtcagcgta
      361 aatgggtagg gggcttcaaa tcgtcccccc atacgatata agttgttact agtgcttgga
      421 ttctcaccaa taaaaaacgc ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt
      481 tctgaggtca ttactggatc tatcaacagg agtccaagcg agctcgatat caaattacgc
      541 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
      601 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
      661 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
      721 taaatcaaaa ctggtgaaac tcacccaggg attggctgag acgaaaaaca tattctcaat
      781 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
      841 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
      901 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
      961 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
     1021 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
     1081 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
     1141 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
     1201 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
     1261 attatggtga aagttggaac ctcttacgtg ccgatcaacg tctcattgat acctgaaaca
     1321 aaacccatcg tacggccaag gaagtctcca ataactgtga tccaccacaa gcgccagggt
     1381 tttcccagtc acgacgttgt aaaacgacgg ccagtcatgc ataatccgca cgcatctgga
     1441 ataaggaagt gccattccgc ctgaccttag cgtgcgaaca gcacgctatt gttgtatcat
     1501 tactcgcatc cattctcagg ctgtctcgtc tcgtctccga ggtctccgga gatcacgtgc
     1561 tataaaaata attataattt aaatttttta atataaatat ataaattaaa aatagaaagt
     1621 aaaaaaagaa attaaagaaa aaatagtttt tgttttccga agatgtaaaa gactctaggg
     1681 ggatcgccaa caaatactac cttttatctt gctcgtcctg ctctcaggta ttaatgccga
     1741 attgtttcat cttgtctgtg tagaagacca cacacgaaaa tcctgtgatt ttacatttta
     1801 cttatcgtta atcgaatgta tatctattta atctgctttt cttgtctaat aaatatatat
     1861 gtaaagtacg ctttttgttg aaatttttta aacctttgtt tatttttttt tcttcattcc
     1921 gtaactcttc taccttcttt atttactttc taaaatccaa atacaaaaca taaaaataaa
     1981 taaacacaga gtaaattccc aaattattcc atcattaaaa gatacgaggc gcgtgtaagt
     2041 tacaggcaag cgatccgctc gagaccaccc caggatacat agattaccac aactccgagc
     2101 ccttccacct agcgtgctaa ccacgcacgc tattgttgta taggctaggt ggaggctcag
     2161 tgatgataag tctgcgatgg tggatgcatg tgtcatggtc atagctgttt cctgtgtgaa
     2221 attgttatcc gctcagaggg cacaatccta ttccgcgcta tccgacaatc tccaagacat
     2281 taggtggagt tcagttcggc gtatggcata tgtcgctgga aagaacatgt gagcaaaagg
     2341 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2401 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     2461 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     2521 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     2581 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     2641 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaact
//
