LOCUS       sequence-402922-        4049 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 4049)
  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 4049)
  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_23 [pSC101ts ORI]
FEATURES             Location/Qualifiers
     source          1..4049
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      565..787
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             835..1785
                     /codon_start=1
                     /gene="rep101 encoding an A56V mutation"
                     /product="temperature-sensitive version of the RepA protein
                     needed for replication with the pSC101 origin (Armstrong et
                     al., 1984)"
                     /label=Rep101(Ts)
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYVQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     primer_bind     complement(2093..2109)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2093..2109)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2106..2128)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(2281..2869)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2361..2380)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(3002..3096)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3117..3776)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKALMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3691..3710
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(3777..3879)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3966..3988
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3980..3997
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3981..3997
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctca ggaggacccc gttgatgata ccgctgcctt actgggtgca
      121 ttagccagtc tgaatgacct gtcacgggat aatccgaagt ggtcagactg gaaaatcaga
      181 gggcaggaac tgctgaacag caaaaagtca gatagcacca catagcagac ccgccataaa
      241 acgccctgag aagcccgtga cgggcttttc ttgtattatg ggtagtttcc ttgcatgaat
      301 ccataaaagg cgcctgtagt gccatttacc cccattcact gccagagccg tgagcgcagc
      361 gaactgaatg tcacgaaaaa gacagcgact caggtgcctg atggtcggag acaaaaggaa
      421 tattcagcga tttgcccgat tgcggccgca accgagcttg cgagggtgct acttaagcct
      481 ttagggtttt aaggtctgtt ttgtagagga gcaaacagcg tttgcgacat ccttttgtaa
      541 tactgcggaa ctgactaaag tagtgagtta tacacagggc tgggatctat tctttttatc
      601 tttttttatt ctttctttat tctataaatt ataaccactt gaatataaac aaaaaaaaca
      661 cacaaaggtc tagcggaatt tacagagggt ctagcagaat ttacaagttt tccagcaaag
      721 gtctagcaga atttacagat acccacaact caaaggaaaa ggactagtaa ttatcattga
      781 ctagcccatc tcaattggta tagtgattaa aatcacctag accaattgag atgtatgtct
      841 gaattagttg ttttcaaagc aaatgaacta gcgattagtc gctatgactt aacggagcat
      901 gaaaccaagc taattttatg ctgtgtggca ctactcaacc ccacgattga aaaccctaca
      961 aggaaagaac ggacggtatc gttcacttat aaccaatacg ttcagatgat gaacatcagt
     1021 agggaaaatg cttatggtgt attagctaaa gcaaccagag agctgatgac gagaactgtg
     1081 gaaatcagga atcctttggt taaaggcttt gagattttcc agtggacaaa ctatgccaag
     1141 ttctcaagcg aaaaattaga attagttttt agtgaagaga tattgcctta tcttttccag
     1201 ttaaaaaaat tcataaaata taatctggaa catgttaagt cttttgaaaa caaatactct
     1261 atgaggattt atgagtggtt attaaaagaa ctaacacaaa agaaaactca caaggcaaat
     1321 atagagatta gccttgatga atttaagttc atgttaatgc ttgaaaataa ctaccatgag
     1381 tttaaaaggc ttaaccaatg ggttttgaaa ccaataagta aagatttaaa cacttacagc
     1441 aatatgaaat tggtggttga taagcgaggc cgcccgactg atacgttgat tttccaagtt
     1501 gaactagata gacaaatgga tctcgtaacc gaacttgaga acaaccagat aaaaatgaat
     1561 ggtgacaaaa taccaacaac cattacatca gattcctacc tacataacgg actaagaaaa
     1621 acactacacg atgctttaac tgcaaaaatt cagctcacca gttttgaggc aaaatttttg
     1681 agtgacatgc aaagtaagta tgatctcaat ggttcgttct catggctcac gcaaaaacaa
     1741 cgaaccacac tagagaacat actggctaaa tacggaagga tctgaggttc ttatggctct
     1801 tgtatctatc agtgaagcat caagactaac acgcttgaga ccaccccagg atacatagat
     1861 taccacaact ccgagccctt ccacctagcg tgctaaccac gcacgctatt gttgtatccc
     1921 atataatgca atcgtagtct acctgactgt acttagaaat gtggcttcgc ctttgcccac
     1981 gcacctgatc gctcctcgtt tgcttttaag gaccggacga accacagagc attagaaggc
     2041 taggtggagg ctcagtgatg ataagtctgc gatggtggat gcatgtgtca tggtcatagc
     2101 tgtttcctgt gtgaaattgt tatccgctca gagggcacaa tcctattccg cgctatccga
     2161 caatctccaa gacattaggt ggagttcagt tcggcgtatg gcatatgtcg ctggaaagaa
     2221 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     2281 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     2341 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     2401 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     2461 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     2521 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     2581 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     2641 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     2701 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     2761 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     2821 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     2881 gatcttttct acggggtctg acgctctatt caacaaagcc gccgtcccgt caagtcagcg
     2941 taaatgggta gggggcttca aatcgtcccc ccatacgata taagttgtta ctagtgcttg
     3001 gattctcacc aataaaaaac gcccggcggc aaccgagcgt tctgaacaaa tccagatgga
     3061 gttctgaggt cattactgga tctatcaaca ggagtccaag cgagctcgat atcaaattac
     3121 gccccgccct gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg
     3181 aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcgc cttgtcgcct
     3241 tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga agttgtccat attggccacg
     3301 tttaaatcaa aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca
     3361 ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat
     3421 atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca
     3481 gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg
     3541 tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag aatgtgaata
     3601 aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc
     3661 agctgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct
     3721 ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta
     3781 gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt
     3841 tcattatggt gaaagttgga acctcttacg tgccgatcaa cgtctcattg atacctgaaa
     3901 caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac aagcgccagg
     3961 gttttcccag tcacgacgtt gtaaaacgac ggccagtcat gcataatccg cacgcatctg
     4021 gaataaggaa gtgccattcc gcctgacct
//
