LOCUS       sequence-355199-        2850 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Golden Gate GFP dropout vector to generate S. thermophilus type I-E 
            single arrays.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2850)
  AUTHORS   Wimmer F, Mougiakos I, Englert F, Beisel CL
  TITLE     Rapid cell-free characterization of multi-subunit CRISPR effectors 
            and transposons.
  JOURNAL   Mol Cell. 2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026.
  PUBMED    35216669
REFERENCE   2  (bases 1 to 2850)
  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: "Mol Cell. 
            2022 Feb 16. pii: S1097-2765(22)00102-2. doi: 
            10.1016/j.molcel.2022.01.026."
COMMENT     Sequence Label: Sth_I-E array_GFP dropout
FEATURES             Location/Qualifiers
     source          1..2850
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     569..588
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      836..1424
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1325..1344
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1546..1580
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     RBS             1767..1778
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1785..2498
                     /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"
     terminator      2510..2581
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2597..2624
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(2740..2774)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(join(2845..2850,1..654))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 cccgccctgc cactcatcgc agtactgttg taattcatta agcattctgc cgacatggaa
       61 gccatcacaa acggcatgat gaacctgaat cgccagcggc atcagcacct tgtcgccttg
      121 cgtataatat ttgcccatgg tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt
      181 taaatcaaaa ctggtgaaac tcacccaggg attggctgaa acgaaaaaca tattctcaat
      241 aaacccttta gggaaatagg ccaggttttc accgtaacac gccacatctt gcgaatatat
      301 gtgtagaaac tgccggaaat cgtcgtggta ttcactccag agcgatgaaa acgtttcagt
      361 ttgctcatgg aaaacggtgt aacaagggtg aacactatcc catatcacca gctcaccgtc
      421 tttcattgcc atacgaaatt ccggatgagc attcatcagg cgggcaagaa tgtgaataaa
      481 ggccggataa aacttgtgct tatttttctt tacggtcttt aaaaaggccg taatatccag
      541 ctgaacggtc tggttatagg tacattgagc aactgactga aatgcctcaa aatgttcttt
      601 acgatgccat tgggatatat caacggtggt atatccagtg atttttttct ccattttagc
      661 ttccttagct cctgaaaatc tcgataactc aaaaaatacg cccggtagtg atcttatttc
      721 attatggtga aagttggaac ctcttacgtg cccgatcaat catgaccaaa atcccttaac
      781 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
      841 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
      901 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
      961 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     1021 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     1081 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     1141 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     1201 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     1261 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     1321 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     1381 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     1441 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     1501 cccctgattc tgtggataac cgtcgagagc atcgatccta gcatgttgac agctagctca
     1561 gtcctaggta taatgctagc gtttttcccg cacacgcggg ggtgatccgg gtcttcgtga
     1621 aagtgaaacg tgatttcatg cgtcattttg aacattttgt aaatcttatt taataatgtg
     1681 tgcggcaatt cacatttaat ttatgaatgt tttcttaaca tcgcggcaac tcaagaaacg
     1741 gcaggttcgg atcttagcta ctagagaaag aggagaaata ctagatgcgt aaaggcgaag
     1801 agctgttcac tggtgtcgtc cctattctgg tggaactgga tggtgatgtc aacggtcata
     1861 agttttccgt gcgtggcgag ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt
     1921 tcatctgtac tactggtaaa ctgccggttc cttggccgac tctggtaacg acgctgactt
     1981 atggtgttca gtgctttgct cgttatccgg accatatgaa gcagcatgac ttcttcaagt
     2041 ccgccatgcc ggaaggctat gtgcaggaac gcacgatttc ctttaaggat gacggcacgt
     2101 acaaaacgcg tgcggaagtg aaatttgaag gcgataccct ggtaaaccgc attgagctga
     2161 aaggcattga ctttaaagag gacggcaata tcctgggcca taagctggaa tacaatttta
     2221 acagccacaa tgtttacatc accgccgata aacaaaaaaa tggcattaaa gcgaatttta
     2281 aaattcgcca caacgtggag gatggcagcg tgcagctggc tgatcactac cagcaaaaca
     2341 ctccaatcgg tgatggtcct gttctgctgc cagacaatca ctatctgagc acgcaaagcg
     2401 ttctgtctaa agatccgaac gagaaacgcg atcatatggt tctgctggag ttcgtaaccg
     2461 cagcgggcat cacgcatggt atggatgaac tgtacaaatg accaggcatc aaataaaacg
     2521 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     2581 ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata gtgaagacct
     2641 gtttttcccg cacacgcggg ggtgatccaa aaaaaaaccc cgcccctgac agggcggggt
     2701 tttttttaaa gaatcgatcg atcccgaatg agtagaccaa taaaaaacgc ccggcggcaa
     2761 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
     2821 agtccaagcg agctcgatat caaattacgc
//
