LOCUS       sequence-398505-        3014 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Even assembly vector with sfGFP dropout for Even level assemblies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3014)
  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 3014)
  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: pGLDV_20 [31e1G]
FEATURES             Location/Qualifiers
     source          1..3014
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     175..194
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(261..363)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     450..472
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     464..481
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     465..481
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     RBS             778..789
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             796..1509
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1757..1773)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1757..1773)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1770..1792)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      1963..2351
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     terminator      complement(2500..2594)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2615..3014,1..260))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 ctatcccata tcaccagctc accgtctttc attgccatac gaaattccgg atgagcattc
       61 atcaggcggg caagaatgtg aataaaggcc ggataaaact tgtgcttatt tttctttacg
      121 gtctttaaaa aggccgtaat atccagctga acggtctggt tataggtaca ttgagcaact
      181 gactgaaatg cctcaaaatg ttctttacga tgccattggg atatatcaac ggtggtatat
      241 ccagtgattt ttttctccat tttagcttcc ttagctcctg aaaatctcga taactcaaaa
      301 aatacgcccg gtagtgatct tatttcatta tggtgaaagt tggaacctct tacgtgccga
      361 tcaacgtctc attgatacct gaaacaaaac ccatcgtacg gccaaggaag tctccaataa
      421 ctgtgatcca ccacaagcgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
      481 tcatgcataa tccgcacgca tctggaataa ggaagtgcca ttccgcctga ccttagcgtg
      541 cgaacagcac gctattgttg tatcattact cgcatccatt ctcaggctgt ctcgtctcgt
      601 ctccgaggtc tcaggagaat gtgaagagcg aaagtgaaac gtgatttcat gcgtcatttt
      661 gaacattttg taaatcttat ttaataatgt gtgcggcaat tcacatttaa tttatgaatg
      721 ttttcttaac atcgcggcaa ctcaagaaac ggcaggttcg gatcttagct actagagaaa
      781 gaggagaaat actagatgcg taaaggcgaa gaactgttca ctggtgtcgt ccctattctg
      841 gtggaactgg atggtgatgt caacggtcat aagttttccg tgcgtggcga gggtgaaggt
      901 gacgcaacta atggtaaact gacgctgaag ttcatctgta ctactggtaa actgccggtt
      961 ccttggccga ctctggtaac gacgctgact tatggtgttc agtgctttgc tcgttatccg
     1021 gaccatatga agcagcatga cttcttcaag tccgccatgc cggaaggcta tgtgcaggaa
     1081 cgcacgattt cctttaagga tgacggcacg tacaaaacgc gtgcggaagt gaaatttgaa
     1141 ggcgataccc tggtaaaccg cattgagctg aaaggcattg actttaaaga ggacggcaat
     1201 atcctgggcc ataagctgga atacaatttt aacagccaca atgtttacat caccgccgat
     1261 aaacaaaaaa atggcattaa agcgaatttt aaaattcgcc acaacgtgga ggatggcagc
     1321 gtgcagctgg ctgatcacta ccagcaaaac actccaatcg gtgatggtcc tgttctgctg
     1381 ccagacaatc actatctgag cacgcaaagc gttctgtcta aagatccgaa cgagaaacgc
     1441 gatcatatgg ttctgctgga gttcgtaacc gcagcgggca tcacgcatgg tatggatgaa
     1501 ctgtacaaat gattcagcca aaaaacttaa gaccgccggt cttgtccact accttgcagt
     1561 aatgcggtgg acaggatcgg cggttttctt ttctcttctc aagctcttca ggtatacttg
     1621 agaccacccc aggatacata gattaccaca actccgagcc cttccaccta gcgtgctaac
     1681 cacgcacgct attgttgtat aggctaggtg gaggctcagt gatgataagt ctgcgatggt
     1741 ggatgcatgt gtcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcagagggc
     1801 acaatcctat tccgcgctat ccgacaatct ccaagacatt aggtggagtt cagttcggcg
     1861 tatggcatat gtcgctggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     1921 taaaaaggcc gcgttgctgg cgttacagct gggcgcgccc catgtcagcc gttaagtgtt
     1981 cctgtgtcac tcaaaattgc tttgagaggc tctaagggct tctcagtgcg ttacatccct
     2041 ggcttgttgt ccacaaccgt taaaccttaa aggctttaaa agccttatat attctttttt
     2101 ttcttataaa acttaaaacc ttagaggcta tttaagttgc tgatttatat taattttatt
     2161 gttcaaacat gagagcttag tacgtgaaac atgagagctt agtacgttag ccatgagagc
     2221 ttagtacgtt agccatgagg gtttagttcg ttaaacatga gagcttagta cgttaaacat
     2281 gagagcttag tacgtgaaac atgagagctt agtacgtact atcaacaggt tgaactgctg
     2341 atcttcagat cggcgcgccg gccggccgaa gatcctttga tcttttctac ggggtctgac
     2401 gctctattca acaaagccgc cgtcccgtca agtcagcgta aatgggtagg gggcttcaaa
     2461 tcgtcccccc atacgatata agttgttact agtgcttgga ttctcaccaa taaaaaacgc
     2521 ccggcggcaa ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc
     2581 tatcaacagg agtccaagcg agctcgatat caaattacgc cccgccctgc cactcatcgc
     2641 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     2701 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     2761 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     2821 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     2881 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     2941 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     3001 aacaagggtg aaca
//
