LOCUS       sequence-397961-        3164 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 0 storage vector with sfGFP dropout for Level 0 parts.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3164)
  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 3164)
  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_17 [380G]
FEATURES             Location/Qualifiers
     source          1..3164
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             237..248
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             255..968
                     /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(1208..1224)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1208..1224)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1221..1243)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1396..1984)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1476..1495)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2117..2211)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(2232..2891)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2806..2825
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2892..2994)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     3081..3103
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3095..3112
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3096..3112
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 tagcgtgcga acagcacgct attgttgtat cattactcgc atccattctc aggctgtctc
       61 gtctcgtctc cgaggtctcc agaagagcga aagtgaaacg tgatttcatg cgtcattttg
      121 aacattttgt aaatcttatt taataatgtg tgcggcaatt cacatttaat ttatgaatgt
      181 tttcttaaca tcgcggcaac tcaagaaacg gcaggttcgg atcttagcta ctagagaaag
      241 aggagaaata ctagatgcgt aaaggcgaag aactgttcac tggtgtcgtc cctattctgg
      301 tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag ggtgaaggtg
      361 acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa ctgccggttc
      421 cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct cgttatccgg
      481 accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat gtgcaggaac
      541 gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg aaatttgaag
      601 gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagag gacggcaata
      661 tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc accgccgata
      721 aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag gatggcagcg
      781 tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct gttctgctgc
      841 cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac gagaaacgcg
      901 atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt atggatgaac
      961 tgtacaaatg attcagccaa aaaacttaag accgccggtc ttgtccacta ccttgcagta
     1021 atgcggtgga caggatcggc ggttttcttt tctcttctca agctcttcac gagaccaccc
     1081 caggatacat agattaccac aactccgagc ccttccacct agcgtgctaa ccacgcacgc
     1141 tattgttgta taggctaggt ggaggctcag tgatgataag tctgcgatgg tggatgcatg
     1201 tgtcatggtc atagctgttt cctgtgtgaa attgttatcc gctcagaggg cacaatccta
     1261 ttccgcgcta tccgacaatc tccaagacat taggtggagt tcagttcggc gtatggcata
     1321 tgtcgctgga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     1381 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     1441 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     1501 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     1561 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     1621 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     1681 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     1741 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     1801 cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct
     1861 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     1921 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     1981 tcaagaagat cctttgatct tttctacggg gtctgacgct ctattcaaca aagccgccgt
     2041 cccgtcaagt cagcgtaaat gggtaggggg cttcaaatcg tccccccata cgatataagt
     2101 tgttactagt gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga
     2161 acaaatccag atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc
     2221 tcgatatcaa attacgcccc gccctgccac tcatcgcagt actgttgtaa ttcattaagc
     2281 attctgccga catggaagcc atcacaaacg gcatgatgaa cctgaatcgc cagcggcatc
     2341 agcaccttgt cgccttgcgt ataatatttg cccatggtga aaacgggggc gaagaagttg
     2401 tccatattgg ccacgtttaa atcaaaactg gtgaaactca cccagggatt ggctgagacg
     2461 aaaaacatat tctcaataaa ccctttaggg aaataggcca ggttttcacc gtaacacgcc
     2521 acatcttgcg aatatatgtg tagaaactgc cggaaatcgt cgtggtattc actccagagc
     2581 gatgaaaacg tttcagtttg ctcatggaaa acggtgtaac aagggtgaac actatcccat
     2641 atcaccagct caccgtcttt cattgccata cgaaattccg gatgagcatt catcaggcgg
     2701 gcaagaatgt gaataaaggc cggataaaac ttgtgcttat ttttctttac ggtctttaaa
     2761 aaggccgtaa tatccagctg aacggtctgg ttataggtac attgagcaac tgactgaaat
     2821 gcctcaaaat gttctttacg atgccattgg gatatatcaa cggtggtata tccagtgatt
     2881 tttttctcca ttttagcttc cttagctcct gaaaatctcg ataactcaaa aaatacgccc
     2941 ggtagtgatc ttatttcatt atggtgaaag ttggaacctc ttacgtgccg atcaacgtct
     3001 cattgatacc tgaaacaaaa cccatcgtac ggccaaggaa gtctccaata actgtgatcc
     3061 accacaagcg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtcatgcata
     3121 atccgcacgc atctggaata aggaagtgcc attccgcctg acct
//
