LOCUS       sequence-401236-        2834 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 2834)
  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 2834)
  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_65 [StayGold]
FEATURES             Location/Qualifiers
     source          1..2834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(57..73)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(57..73)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(70..92)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(245..833)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(325..344)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(966..1060)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1081..1740)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     1655..1674
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(1741..1843)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     1930..1952
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1944..1961
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1945..1961
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             2095..2745
                     /codon_start=1
                     /product="bright and highly photostable dimeric green 
                     fluorescent protein (Hirano et al., 2022)"
                     /label=StayGold
                     /note="engineered variant of a GFP from Cytaeis uchidae"
                     /translation="MASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKGKYVCTSGKL
                     PMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQYKGDGSIHAKHQHFM
                     KNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVQHIPRDDGVECPVTLLYPLLSDKS
                     KCVEAHQNTICKPLHNQPAPDVPYHWIRKQYTQSKDDTEERDHICQSETLEAHL"
ORIGIN
        1 aggctaggtg gaggctcagt gatgataagt ctgcgatggt ggatgcatgt gtcatggtca
       61 tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc acaatcctat tccgcgctat
      121 ccgacaatct ccaagacatt aggtggagtt cagttcggcg tatggcatat gtcgctggaa
      181 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
      241 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
      301 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
      361 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
      421 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
      481 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
      541 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
      601 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
      661 ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag
      721 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
      781 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
      841 ctttgatctt ttctacgggg tctgacgctc tattcaacaa agccgccgtc ccgtcaagtc
      901 agcgtaaatg ggtagggggc ttcaaatcgt ccccccatac gatataagtt gttactagtg
      961 cttggattct caccaataaa aaacgcccgg cggcaaccga gcgttctgaa caaatccaga
     1021 tggagttctg aggtcattac tggatctatc aacaggagtc caagcgagct cgatatcaaa
     1081 ttacgccccg ccctgccact catcgcagta ctgttgtaat tcattaagca ttctgccgac
     1141 atggaagcca tcacaaacgg catgatgaac ctgaatcgcc agcggcatca gcaccttgtc
     1201 gccttgcgta taatatttgc ccatggtgaa aacgggggcg aagaagttgt ccatattggc
     1261 cacgtttaaa tcaaaactgg tgaaactcac ccagggattg gctgagacga aaaacatatt
     1321 ctcaataaac cctttaggga aataggccag gttttcaccg taacacgcca catcttgcga
     1381 atatatgtgt agaaactgcc ggaaatcgtc gtggtattca ctccagagcg atgaaaacgt
     1441 ttcagtttgc tcatggaaaa cggtgtaaca agggtgaaca ctatcccata tcaccagctc
     1501 accgtctttc attgccatac gaaattccgg atgagcattc atcaggcggg caagaatgtg
     1561 aataaaggcc ggataaaact tgtgcttatt tttctttacg gtctttaaaa aggccgtaat
     1621 atccagctga acggtctggt tataggtaca ttgagcaact gactgaaatg cctcaaaatg
     1681 ttctttacga tgccattggg atatatcaac ggtggtatat ccagtgattt ttttctccat
     1741 tttagcttcc ttagctcctg aaaatctcga taactcaaaa aatacgcccg gtagtgatct
     1801 tatttcatta tggtgaaagt tggaacctct tacgtgccga tcaacgtctc attgatacct
     1861 gaaacaaaac ccatcgtacg gccaaggaag tctccaataa ctgtgatcca ccacaagcgc
     1921 cagggttttc ccagtcacga cgttgtaaaa cgacggccag tcatgcataa tccgcacgca
     1981 tctggaataa ggaagtgcca ttccgcctga ccttagcgtg cgaacagcac gctattgttg
     2041 tatcattact cgcatccatt ctcaggctgt ctcgtctcgt ctccgaggtc tcaaatggcc
     2101 tcaaccccgt ttaagtttca gttaaaaggc acgatcaatg gcaagagctt cacagtagaa
     2161 ggtgaaggag aaggtaatag ccacgaaggc agccataaag gcaagtatgt ttgtacgtcc
     2221 ggtaaactgc ctatgagttg ggcggctttg ggcacctcct tcgggtacgg aatgaaatac
     2281 tacacaaagt atccgtcggg tctgaaaaac tggtttcatg aagtgatgcc ggaaggcttc
     2341 acttacgatc gccatattca gtataaaggt gatggcagca tccatgcgaa acatcagcat
     2401 tttatgaaaa atggcaccta tcacaacatt gtagaattca cgggccagga ctttaaagag
     2461 aatagccccg tgttaacggg tgacatgaat gtctcgctgc ccaacgaggt gcaacacatt
     2521 cctcgcgatg acggggttga atgtccagtt accctgttgt acccgcttct atcagataaa
     2581 agtaaatgcg tcgaagccca tcagaacacc atttgcaagc cgctgcacaa ccaaccggcg
     2641 ccagatgtgc cgtatcactg gattcgtaaa caatataccc agtctaaaga tgataccgaa
     2701 gagagggacc atatctgcca gtctgagact ctcgaggcac acctgtaagc tttgagacca
     2761 ccccaggata catagattac cacaactccg agcccttcca cctagcgtgc taaccacgca
     2821 cgctattgtt gtat
//
