LOCUS       sequence-406394-        7483 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 7483)
  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 7483)
  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_18 [AMA1 long ORI]
FEATURES             Location/Qualifiers
     source          1..7483
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(223..317)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(338..997)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     912..931
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(998..1100)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1187..1209
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1201..1218
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1202..1218
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      1363..6611
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     primer_bind     complement(6797..6813)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6797..6813)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6810..6832)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(join(6985..7483,1..90))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7065..7084)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
       61 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
      121 gacgctctat tcaacaaagc cgccgtcccg tcaagtcagc gtaaatgggt agggggcttc
      181 aaatcgtccc cccatacgat ataagttgtt actagtgctt ggattctcac caataaaaaa
      241 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
      301 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
      361 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
      421 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
      481 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
      541 aactcaccca gggattggct gagacgaaaa acatattctc aataaaccct ttagggaaat
      601 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
      661 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
      721 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
      781 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
      841 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
      901 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
      961 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     1021 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     1081 aacctcttac gtgccgatca acgtctcatt gatacctgaa acaaaaccca tcgtacggcc
     1141 aaggaagtct ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt
     1201 tgtaaaacga cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc
     1261 cgcctgacct tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc
     1321 aggctgtctc gtctcgtctc cgaggtctcc ggagagcttg atagcttatt ttttgtatac
     1381 tgttttgtga tagcacgaag tttttccacg gtatcttgtt aaaaatatat atttgtggcg
     1441 ggcttaccta catcaaatta ataagagact aattataaac taaacacaca agcaagctac
     1501 tttagggtaa aagtttataa atgcttttga cgtataaacg ttgcttgtat ttattattac
     1561 aattaaaggt ggatagaaaa cctagagact agttagaaac taatctcagg tttgcgttaa
     1621 actaaatcag agcccgagag gttaacagaa cctagaaggg gactagatat ccgggtaggg
     1681 aaacaaaaaa aaaaaacaag acagccacat attagggaga ctagttagaa gctagttcca
     1741 ggactaggaa aataaaagac aatgatacca cagtctagtt gacaactaga tagattctag
     1801 attgaggcca aagtctctga gatccaggtt agttgcaact aatactagtt agtatctagt
     1861 ctcctataac tctgaagcta gaataactta ctactattat cctcaccact gttcagctgc
     1921 gcaaacggag tgattgcaag gtgttcagag actagttatt gactagtcag tgactagcaa
     1981 taactaacaa ggtattaacc taccatgtct gccatcaccc tgcacttcct cgggctcagc
     2041 agccttttcc tcctcatttt catgctcatt ttccttgttt aagactgtga ctagtcaaag
     2101 actagtccag aaccacaaag gagaaatgtc ttaccacttt cttcattgct tgtctctttt
     2161 gcattatcca tgtctgcaac tagttagagt ctagttagtg actagtccga cgaggacttg
     2221 cttgtctccg gattgttgga ggaactctcc agggcctcaa gatccacaac agagccttct
     2281 agaagactgg tcaataacta gttggtcttt gtctgagtct gacttacgag gttgcatact
     2341 cgctcccttt gcctcgtcaa tcgatgagaa aaagcgccaa aactcgcaat atggctttga
     2401 accacacggt gctgagacta gttagaatct agtcccaaac tagcttggat agcttacctt
     2461 tgccctttgc gttgcgacag gtcttgcagg gtatggttcc tttctcacca gctgatttag
     2521 ctgccttgct accctcacgg cggatctgca taaagagtgg ctagaggtta taaattagca
     2581 ctgatcctag gtacggggct gaatgtaact tgcctttcct ttctcatcgc gcggcaagac
     2641 aggcttgctc aaattcctac cagtcacagg ggtatgcacg gcgtacggac cacttgaact
     2701 agtcacagat tagttagcaa ctagtctgca ttgaatggct gtacttacgg gccctcgcca
     2761 ttgtcctgat catttccagc ttcaccctcg ttgctgcaaa gtagttagtg actagtcaag
     2821 gactagttga aatgggagaa gaaactcacg aattctcgac acccttagta ttgtggtcct
     2881 tggacttggt gctgctatat attagctaat acactagtta gactcacaga aacttacgca
     2941 gctcgcttgc gcttcttggt aggagtcggg gttgggagaa cagtgccttc aaacaagcct
     3001 tcataccatg ctacttgact agtcagggac tagtcaccaa gtaatctaga taggacttgc
     3061 ctttggcctc catcagttcc ttcatagtgg gaggtccatt gtgcaatgta aactccatgc
     3121 cgtgggagtt cttgtccttc aagtgcttga ccaatatgtt tctgttggca gagggaacct
     3181 gtcaactagt taataactag tcagaaacta gtatagcagt agactcactg tacgcttgag
     3241 gcatcccttc actcggcagt agacttcata tggatggata tcaggcacgc cattgtcgtc
     3301 ctgtggacta gtcagtaact aggcttaaag ctagtcgggt cggcttacta tcttgaaatc
     3361 cggcagcgta agctccccgt ccttaactgc ctcgagatag tgacagtact ctggggactt
     3421 tcggagatcg ttatcgcgaa tgctcggcat actaatcgtt gactagtctt ggactagtcc
     3481 cgagcaaaaa ggattggagg aggaggagga aggtgagagt gagacaaaga gcgaaataag
     3541 agcttcaaag gctatctcta agcagtatga aggttaagta tctagttctt gactagattt
     3601 aaaagagatt tcgactagtt atgtacctgg agtttggata taggaatgtg ttgtggtaac
     3661 gaaatgtaag ggggaggaaa gaaaaagtcg gtcaagaggt aactctaagt cggccattcc
     3721 tttttgggag gcgctaacca taaacggcat ggtcgactta gagttagctc agggaattta
     3781 gggagttatc tgcgaccacc gaggaacggc ggaatgccaa agaatcccga tggagctcta
     3841 gctggcggtt gacaacccca ccttttggcg tttctgcggc gttgcaggcg ggactggata
     3901 cttcgtagaa ccagaaaggc aaggcagaac gcgctcagca agagtgttgg aagtgatagc
     3961 atgatgtgcc ttgttaacta ggtcaaaatc tgcagtatgc ttgatgttat ccaaagtgtg
     4021 agagaggaag gtccaaacat acacgattgg gagagggcct aggtataaga gtttttgagt
     4081 agaacgcatg tgagcccagc catctcgagg agattaaaca cgggccggca tttgatggct
     4141 atgttagtac cccaatggaa agcctgagag tccagtggtc gcagataact ccctaaattc
     4201 cctgagctaa ctctaagtcg accatgccgt ttatggttag cgcctcccaa aaaggaatgg
     4261 ccgacttaga gttacctctt gaccgacttt ttctttcctc ccccttacat ttcgttacca
     4321 caacacattc ctatatccaa actccaggta cataactagt cgaaatctct tttaaatcta
     4381 gtcaagaact agatacttaa ccttcatact gcttagagat agcctttgaa gctcttattt
     4441 cgctctttgt ctcactctca ccttcctcct cctcctccaa tcctttttgc tcgggactag
     4501 tccaagacta gtcaacgatt agtatgccga gcattcgcga taacgatctc cgaaagtccc
     4561 cagagtactg tcactatctc gaggcagtta aggacgggga gcttacgctg ccggatttca
     4621 agatagtaag ccgacccgac tagctttaag cctagttact gactagtcca caggacgaca
     4681 atggcgtgcc tgatatccat ccatatgaag tctactgccg agtgaaggga tgcctcaagc
     4741 gtacagtgag tctactgcta tactagtttc tgactagtta ttaactagtt gacaggttcc
     4801 ctctgccaac agaaacatat tggtcaagca cttgaaggac aagaactccc acggcatgga
     4861 gtttacattg cacaatggac ctcccactat gaaggaactg atggaggcca aaggcaagtc
     4921 ctatctagat tacttggtga ctagtccctg actagtcaag tagcatggta tgaaggcttg
     4981 tttgaaggca ctgttctccc aaccccgact cctaccaaga agcgcaagcg agctgcgtaa
     5041 gtttctgtga gtctaactag tgtattagct aatatatagc agcaccaagt ccaaggacca
     5101 caatactaag ggtgtcgaga attcgtgagt ttcttctccc atttcaacta gtccttgact
     5161 agtcactaac tactttgcag caacgagggt gaagctggaa atgatcagga caatggcgag
     5221 ggcccgtaag tacagccatt caatgcagac tagttgctaa ctaatctgtg actagttcaa
     5281 gtggtccgta cgccgtgcat acccctgtga ctggtaggaa tttgagcaag cctgtcttgc
     5341 cgcgcgatga gaaaggaaag gcaagttaca ttcagccccg tacctaggat cagtgctaat
     5401 ttataacctc tagccactct ttatgcagat ccgccgtgag ggtagcaagg cagctaaatc
     5461 agctggtgag aaaggaacca taccctgcaa gacctgtcgc aacgcaaagg gcaaaggtaa
     5521 gctatccaag ctagtttggg actagattct aactagtctc agcaccgtgt ggttcaaagc
     5581 catattgcga gttttggcgc tttttctcat cgattgacga ggcaaaggga gcgagtatgc
     5641 aacctcgtaa gtcagactca gacaaagacc aactagttat tgaccagtct tctagaaggc
     5701 tctgttgtgg atcttgaggc cctggagagt tcctccaaca atccggagac aagcaagtcc
     5761 tcgtcggact agtcactaac tagactctaa ctagttgcag acatggataa tgcaaaagag
     5821 acaagcaatg aagaaagtgg taagacattt ctcctttgtg gttctggact agtctttgac
     5881 tagtcacagt cttaaacaag gaaaatgagc atgaaaatga ggaggaaaag gctgctgagc
     5941 ccgaggaagt gcagggtgat ggcagacatg gtaggttaat accttgttag ttattgctag
     6001 tcactgacta gtcaataact agtctctgaa caccttgcaa tcactccgtt tgcgcagctg
     6061 aacagtggtg aggataatag tagtaagtta ttctagcttc agagttatag gagactagat
     6121 actaactagt attagttgca actaacctgg atctcagaga ctttggcctc aatctagaat
     6181 ctatctagtt gtcaactaga ctgtggtatc attgtctttt attttcctag tcctggaact
     6241 agcttctaac tagtctccct aatatgtggc tgtcttgttt tttttttttg tttccctacc
     6301 cggatatcta gtccccttct aggttctgtt aacctctcgg gctctgattt agtttaacgc
     6361 aaacctgaga ttagtttcta actagtctct aggttttcta tccaccttta attgtaataa
     6421 taaatacaag caacgtttat acgtcaaaag catttataaa cttttaccct aaagtagctt
     6481 gcttgtgtgt ttagtttata attagtctct tattaatttg atgtaggtaa gcccgccaca
     6541 aatatatatt tttaacaaga taccgtggaa aaacttcgtg ctatcacaaa acagtataca
     6601 aaaaataagc tatcgaattc ctgcagcccc agatcatcct gtcttcagtc ttaacgctcg
     6661 agaccacccc aggatacata gattaccaca actccgagcc cttccaccta gcgtgctaac
     6721 cacgcacgct attgttgtat aggctaggtg gaggctcagt gatgataagt ctgcgatggt
     6781 ggatgcatgt gtcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc
     6841 acaatcctat tccgcgctat ccgacaatct ccaagacatt aggtggagtt cagttcggcg
     6901 tatggcatat gtcgctggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     6961 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     7021 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     7081 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     7141 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     7201 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     7261 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     7321 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     7381 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     7441 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggt
//
