LOCUS       sequence-401228-        3676 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 3676)
  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 3676)
  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_58 [tetL]
FEATURES             Location/Qualifiers
     source          1..3676
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             82..1458
                     /codon_start=1
                     /gene="Bacillus subtilis tetL"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MNTSYSQSNLRHNQILIWLCILSFFSVLNEMVLNVSLPDIANDFN
                     KPPASTNWVNTAFMLTFSIGTAVYGKLSDQLGIKRLLLFGIIINCFGSVIGFVGHSFFS
                     LLIMARFIQGAGAAAFPALVMVVVARYIPKENRGKAFGLIGSIVAMGEGVGPAIGGMIA
                     HYIHWSYLLLIPMITIITVPFLMKLLKKEVRIKGHFDIKGIILMSVGIVFFMLFTTSYS
                     ISFLIVSVLSFLIFVKHIRKVTDPFVDPGLGKNIPFMIGVLCGGIIFGTVAGFVSMVPY
                     MMKDVHQLSTAEIGSVIIFPGTMSVIIFGYIGGILVDRRGPLYVLNIGVTFLSVSFLTA
                     SFLLETTSWFMTIIIVFVLGGLSFTKTVISTIVSSSLKQQEAGAGMSLLNFTSFLSEGT
                     GIAIVGGLLSIPLLDQRLLPMEVDQSTYLYSNLLLLFSGIIVISWLVTLNVYKHSQRDF
                     "
     primer_bind     complement(1720..1736)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1720..1736)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1733..1755)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1908..2496)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1988..2007)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2629..2723)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2744..3403)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3318..3337
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3404..3506)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3593..3615
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3607..3624
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3608..3624
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca aatgaataca tcctattcac aatcgaattt acgacacaac
      121 caaattttaa tttggctttg cattttatct ttttttagcg tattaaatga aatggttttg
      181 aacgtcagct tacctgatat tgcaaatgat tttaataaac ctcctgcgag tacaaactgg
      241 gtgaacacag cctttatgtt aaccttttcc attggaacag ctgtatatgg aaagctatct
      301 gatcaattag gcatcaaaag gttactccta tttggaatta taataaattg tttcgggtcg
      361 gtaattgggt ttgttggcca ttctttcttt tccttactta ttatggctcg ttttattcaa
      421 ggggctggtg cagctgcatt tccagcactc gtaatggttg tagttgcgcg ctatattcca
      481 aaggaaaata ggggtaaagc atttggtctt attggatcga tagtagccat gggagaagga
      541 gtcggtccag cgattggtgg aatgatagcc cattatattc attggtccta tcttctactc
      601 attcctatga taacaattat cactgttccg tttcttatga aattattaaa gaaagaagta
      661 aggataaaag gtcattttga tatcaaagga attatactaa tgtctgtagg cattgtattt
      721 tttatgttgt ttacaacatc atatagcatt tcttttctta tcgttagcgt gctgtcattc
      781 ctgatatttg taaaacatat caggaaagta acagatcctt ttgttgatcc cggattaggg
      841 aaaaatatac cttttatgat tggagttctt tgtgggggaa ttatatttgg aacagtagca
      901 gggtttgtct ctatggttcc ttatatgatg aaagatgttc accagctaag tactgccgaa
      961 atcggaagtg taattatttt ccctggaaca atgagtgtca ttattttcgg ctacattggt
     1021 gggatacttg ttgatagaag aggtccttta tacgtgttaa acatcggagt tacatttctt
     1081 tctgttagct ttttaactgc ttcctttctt ttagaaacaa catcatggtt catgacaatt
     1141 ataatcgtat ttgttttagg tgggctttcg ttcaccaaaa cagttatatc aacaattgtt
     1201 tcaagtagct tgaaacagca ggaagctggt gctggaatga gtttgcttaa ctttaccagc
     1261 tttttatcag agggaacagg tattgcaatt gtaggtggtt tattatccat acccttactt
     1321 gatcaaaggt tgttacctat ggaagttgat cagtcaactt atctgtatag taatttgtta
     1381 ttactttttt caggaatcat tgtcattagt tggctggtta ccttgaatgt atataaacat
     1441 tctcaaaggg atttctaagc tttgagacca ccccaggata catagattac cacaactccg
     1501 agcccttcca cctagcgtgc taaccacgca cgctattgtt gtatcccata taatgcaatc
     1561 gtagtctacc tgactgtact tagaaatgtg gcttcgcctt tgcccacgca cctgatcgct
     1621 cctcgtttgc ttttaaggac cggacgaacc acagagcatt agaaggctag gtggaggctc
     1681 agtgatgata agtctgcgat ggtggatgca tgtgtcatgg tcatagctgt ttcctgtgtg
     1741 aaattgttat ccgctcagag ggcacaatcc tattccgcgc tatccgacaa tctccaagac
     1801 attaggtgga gttcagttcg gcgtatggca tatgtcgctg gaaagaacat gtgagcaaaa
     1861 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     1921 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     1981 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2041 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2101 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2161 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     2221 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     2281 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     2341 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     2401 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     2461 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     2521 gggtctgacg ctctattcaa caaagccgcc gtcccgtcaa gtcagcgtaa atgggtaggg
     2581 ggcttcaaat cgtcccccca tacgatataa gttgttacta gtgcttggat tctcaccaat
     2641 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     2701 tactggatct atcaacagga gtccaagcga gctcgatatc aaattacgcc ccgccctgcc
     2761 actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag ccatcacaaa
     2821 cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc gtataatatt
     2881 tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt aaatcaaaac
     2941 tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata aaccctttag
     3001 ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg tgtagaaact
     3061 gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt tgctcatgga
     3121 aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct ttcattgcca
     3181 tacgaaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag gccggataaa
     3241 acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc tgaacggtct
     3301 ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta cgatgccatt
     3361 gggatatatc aacggtggta tatccagtga tttttttctc cattttagct tccttagctc
     3421 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     3481 agttggaacc tcttacgtgc cgatcaacgt ctcattgata cctgaaacaa aacccatcgt
     3541 acggccaagg aagtctccaa taactgtgat ccaccacaag cgccagggtt ttcccagtca
     3601 cgacgttgta aaacgacggc cagtcatgca taatccgcac gcatctggaa taaggaagtg
     3661 ccattccgcc tgacct
//
