LOCUS       sequence-110210-        3108 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes sgRNA Dropout as a Type 234 part to be used in the Dueber 
            YTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3108)
  AUTHORS   Lee ME, DeLoache WC, Cervantes B, Dueber JE
  TITLE     A Highly Characterized Yeast Toolkit for Modular, Multipart Assembly
  JOURNAL   ACS Synth Biol. 2015 Apr 14.
  PUBMED    25871405
REFERENCE   2  (bases 1 to 3108)
  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: "ACS Synth 
            Biol. 2015 Apr 14."
COMMENT     Sequence Label: pYTK050
FEATURES             Location/Qualifiers
     source          1..3108
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             430..441
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             448..1161
                     /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      1173..1244
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1260..1287
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_RNA        1301..1376
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     terminator      complement(1473..1507)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1578..2237)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2152..2171
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2419..3007
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2908..2927
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tcggtctcca acgatgtgct tcagtattac attttttgcc ttcaacgcct tgattgttct
       61 atttttgcta ataataaatc tatttcatcg gactaaaagt ccattagttg taagcggatt
      121 tagctcagtt gggagagcgc cagactgaag aaaaacttcg gtcaagtcat ctggaggtcc
      181 tgtgttcgat ccacagaatt cgcagatggc cggcatggtc ccagcctcct cgctggcgcc
      241 ggctgggcaa caccttcggg tggcgaatgg gacttgagac ggaaagtgaa acgtgatttc
      301 atgcgtcatt ttgaacattt tgtaaatctt atttaataat gtgtgcggca attcacattt
      361 aatttatgaa tgttttctta acatcgcggc aactcaagaa acggcaggtt cggatcttag
      421 ctactagaga aagaggagaa atactagatg cgtaaaggcg aagagctgtt cactggtgtc
      481 gtccctattc tggtggaact ggatggtgat gtcaacggtc ataagttttc cgtgcgtggc
      541 gagggtgaag gtgacgcaac taatggtaaa ctgacgctga agttcatctg tactactggt
      601 aaactgccgg ttccttggcc gactctggta acgacgctga cttatggtgt tcagtgcttt
      661 gctcgttatc cggaccatat gaagcagcat gacttcttca agtccgccat gccggaaggc
      721 tatgtgcagg aacgcacgat ttcctttaag gatgacggca cgtacaaaac gcgtgcggaa
      781 gtgaaatttg aaggcgatac cctggtaaac cgcattgagc tgaaaggcat tgactttaaa
      841 gaggacggca atatcctggg ccataagctg gaatacaatt ttaacagcca caatgtttac
      901 atcaccgccg ataaacaaaa aaatggcatt aaagcgaatt ttaaaattcg ccacaacgtg
      961 gaggatggca gcgtgcagct ggctgatcac taccagcaaa acactccaat cggtgatggt
     1021 cctgttctgc tgccagacaa tcactatctg agcacgcaaa gcgttctgtc taaagatccg
     1081 aacgagaaac gcgatcatat ggttctgctg gagttcgtaa ccgcagcggg catcacgcat
     1141 ggtatggatg aactgtacaa atgaccaggc atcaaataaa acgaaaggct cagtcgaaag
     1201 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctctactag agtcacactg
     1261 gctcaccttc gggtgggcct ttctgcgttt atacgtctca gttttagagc tagaaatagc
     1321 aagttaaaat aaggctagtc cgttatcaac ttgaaaaagt ggcaccgagt cggtgctttt
     1381 tttatttttt gtcactattg ttatgtaaaa tgccacctct gacagtatgg aacgcaaact
     1441 tctgtctagt ggatagctgt gagaccagac caataaaaaa cgcccggcgg caaccgagcg
     1501 ttctgaacaa atccagatgg agttctgagg tcattactgg atctatcaac aggagtccaa
     1561 gcgagctcga tatcaaatta cgccccgccc tgccactcat cgcagtactg ttgtaattca
     1621 ttaagcattc tgccgacatg gaagccatca caaacggcat gatgaacctg aatcgccagc
     1681 ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca tggtgaaaac gggggcgaag
     1741 aagttgtcca tattggccac gtttaaatca aaactggtga aactcaccca gggattggct
     1801 gaaacgaaaa acatattctc aataaaccct ttagggaaat aggccaggtt ttcaccgtaa
     1861 cacgccacat cttgcgaata tatgtgtaga aactgccgga aatcgtcgtg gtattcactc
     1921 cagagcgatg aaaacgtttc agtttgctca tggaaaacgg tgtaacaagg gtgaacacta
     1981 tcccatatca ccagctcacc gtctttcatt gccatacgaa attccggatg agcattcatc
     2041 aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt gcttattttt ctttacggtc
     2101 tttaaaaagg ccgtaatatc cagctgaacg gtctggttat aggtacattg agcaactgac
     2161 tgaaatgcct caaaatgttc tttacgatgc cattgggata tatcaacggt ggtatatcca
     2221 gtgatttttt tctccatttt agcttcctta gctcctgaaa atctcgataa ctcaaaaaat
     2281 acgcccggta gtgatcttat ttcattatgg tgaaagttgg aacctcttac gtgcccgatc
     2341 aatcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2401 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2461 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2521 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2581 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2641 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2701 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2761 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2821 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2881 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2941 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     3001 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     3061 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtag
//
