LOCUS       pCK303.306              2576 bp ds-DNA     circular     09-MAY-2025
DEFINITION  .
KEYWORDS    "Parental Vector" "Origin" "Resistance" "Construction Method"
            "Promoter" "Terminator" "gRNA" "accession:addgene_35279_42946"
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
FEATURES             Location/Qualifiers
     terminator      90..184
                     /label="lambda t0 terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(215..1009)
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     CDS             complement(215..1009)
                     /label="Translation 1009-215"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
     primer_bind     complement(326..345)
                     /label="Neo-F"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     936..955
                     /label="Neo-R"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     misc_feature    1010..1127
                     /label="promoter"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     promoter        1271..1305
                     /label="J23119 (SpeI) promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    1306..1325
                     /label="J306"
                     /ApEinfo_revcolor="#800080"
                     /ApEinfo_fwdcolor="#800080"
     misc_feature    1326..1367
                     /label="JAD designed chimera RNA 3'UTR"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    1372..1386
                     /label="Streptococcus pyogenes tracrRNAterminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1373..1386
                     /label="Hairpin 3"
                     /ApEinfo_revcolor="#b1ff67"
                     /ApEinfo_fwdcolor="#b1ff67"
     misc_feature    1387..1388
                     /label="Left stem extension"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1387..1409
                     /label="MS2 with 2bp stem extension"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1408..1409
                     /label="Right stem extension"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1410..1416
                     /label="SUP4 terminator"
                     /ApEinfo_revcolor="#008080"
                     /ApEinfo_fwdcolor="#008080"
     terminator      1417..1784
                     /label="TrrnB"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     terminator      1710..1777
                     /label="sglTerm"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_fwdcolor="#800040"
     rep_origin      complement(1970..76)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
       61 gggattttgg tcatgagAGT ACTgcttgga ttctcaccaa taaaaaacgc ccggcggcaa
      121 ccgagcgttc tgaacaaatc cagatggagt tctgaggtca ttactggatc tatcaacagg
      181 agtccaagcg agctctcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
      241 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
      301 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
      361 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
      421 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
      481 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
      541 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
      601 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
      661 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
      721 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
      781 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
      841 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
      901 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
      961 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     1021 ctcatcctgt ctcttgatca gatcatgatc ccctgcgcca tcagatcctt ggcggcaaga
     1081 aagccatcca gtttactttg cagggcttcc caaccttacc agagggcgcc ccagctggca
     1141 attccgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1201 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggAATT
     1261 CTAAAGATCT TTGACAGCTA GCTCAGTCCT AGGTATAATA CTAGTTTGTG TCCAGAACGC
     1321 TCCGTGTTTT AGAGCTAGAA ATAGCAAGTT AAAATAAGGC TAGTCCGTTA TCAACTTGAA
     1381 AAAGTGGCAC ATGAGGATCA CCCATGTGCT TTTTTTGAAG CTTGGGCCCG AACAAAAACT
     1441 Catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc attgagttta
     1501 aacggtctcc agcttggctg ttttggcgga tgagagaaga ttttcagcct gatacagatt
     1561 aaatcagaac gcagaagcgg tctgataaaa cagaatttgc ctggcggcag tagcgcggtg
     1621 gtcccacctg accccatgcc gaactcagaa gtgaaacgcc gtagcgccga tggtagtgtg
     1681 gggtctcccc atgcgagagt agggaactgc caggcatcaa ataaaacgaa aggctcagtc
     1741 gaaagactGG GCCTTTCGTT TTATCTGTTG TTTGTCGGTG AACTGGATCC TTACTCGAGT
     1801 CTAGACTGCA Ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     1861 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     1921 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     1981 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     2041 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     2101 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     2161 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     2221 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     2281 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     2341 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     2401 ggtggcctaa ctacggctac actagaagga cagtatttgg tatctgcgct ctgctgaagc
     2461 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     2521 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaa
//