LOCUS       pCK677.102-110          3121 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
     primer_bind     complement(60..79)
                     /label="Neo-F"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     primer_bind     670..689
                     /label="Neo-R"
                     /ApEinfo_revcolor="#faac61"
                     /ApEinfo_fwdcolor="#faac61"
     promoter        1005..1039
                     /label="J23119 (SpeI) promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     None            1040..1059
                     /label="J102"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1060..1172
                     /label="scRNA_2X-BoxB"
                     /ApEinfo_revcolor="#ff9ccd"
                     /ApEinfo_fwdcolor="#ff9ccd"
     misc_feature    1072..1090
                     /label="BoxB"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1127..1145
                     /label="BoxB"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     terminator      1173..1540
                     /label="TrrnB"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     terminator      1466..1533
                     /label="sglTerm"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_fwdcolor="#800040"
     promoter        1550..1584
                     /label="J23119 (SpeI) promoter"
                     /ApEinfo_revcolor="#ffef86"
                     /ApEinfo_fwdcolor="#ffef86"
     misc_feature    1585..1604
                     /label="J110"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1605..1646
                     /label="JAD designed chimera RNA 3'UTR"
                     /ApEinfo_revcolor="#84b0dc"
                     /ApEinfo_fwdcolor="#84b0dc"
     misc_feature    1651..1665
                     /label="Streptococcus pyogenes tracrRNA terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     misc_feature    1666..1667
                     /label="Left stem extension"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1666..1688
                     /label="MS2 with 2bp stem extension"
                     /ApEinfo_revcolor="#f58a5e"
                     /ApEinfo_fwdcolor="#f58a5e"
     misc_feature    1687..1688
                     /label="Right stem extension"
                     /ApEinfo_revcolor="#b4abac"
                     /ApEinfo_fwdcolor="#b4abac"
     misc_feature    1689..1695
                     /label="SUP4 terminator"
                     /ApEinfo_revcolor="#008080"
                     /ApEinfo_fwdcolor="#008080"
     terminator      1696..2063
                     /label="TrrnB"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     terminator      1989..2056
                     /label="sglTerm"
                     /ApEinfo_revcolor="#800040"
                     /ApEinfo_fwdcolor="#800040"
     rep_origin      complement(2249..2931)
                     /label="ColE1 origin"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     terminator      2945..3039
                     /label="lambda t0 terminator"
                     /ApEinfo_revcolor="#85dae9"
                     /ApEinfo_fwdcolor="#85dae9"
     CDS             complement(3070..743)
                     /label="NeoR/KanR"
                     /ApEinfo_revcolor="#c7b0e3"
                     /ApEinfo_fwdcolor="#c7b0e3"
     CDS             complement(3070..743)
                     /label="Translation 743-3070"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRPVLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLSSHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQGLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIALATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
ORIGIN
        1 gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt cgccgccaag ctcttcagca
       61 atatcacggg tagccaacgc tatgtcctga tagcggtccg ccacacccag ccggccacag
      121 tcgatgaatc cagaaaagcg gccattttcc accatgatat tcggcaagca ggcatcgcca
      181 tgggtcacga cgagatcctc gccgtcgggc atgcgcgcct tgagcctggc gaacagttcg
      241 gctggcgcga gcccctgatg ctcttcgtcc agatcatcct gatcgacaag accggcttcc
      301 atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt ggtcgaatgg gcaggtagcc
      361 ggatcaagcg tatgcagccg ccgcattgca tcagccatga tggatacttt ctcggcagga
      421 gcaaggtgag atgacaggag atcctgcccc ggcacttcgc ccaatagcag ccagtccctt
      481 cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa cgcccgtcgt ggccagccac
      541 gatagccgcg ctgcctcgtc ctgcagttca ttcagggcac cggacaggtc ggtcttgaca
      601 aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg cggcatcaga gcagccgatt
      661 gtctgttgtg cccagtcata gccgaatagc ctctccaccc aagcggccgg agaacctgcg
      721 tgcaatccat cttgttcaat catgcgaaac gatcctcatc ctgtctcttg atcagatcat
      781 gatcccctgc gccatcagat ccttggcggc aagaaagcca tccagtttac tttgcagggc
      841 ttcccaacct taccagaggg cgccccagct ggcaattccg acgtctaaga aaccattatt
      901 atcatgacat taacctataa aaataggcgt atcacgaggc agaatttcag ataaaaaaaa
      961 tccttagctt tcgctaagga tgatttctgg AATTCTAAAG ATCTTTGACA GCTAGCTCAG
     1021 TCCTAGGTAT AATACTAGTA GGTATCCGGT GGAACCCAAG TTTGAGAGCT AGGGCCCTGA
     1081 AGAAGGGCCC TAGCAAGTTC AAATAAGGCT AGTCCGTTAT CAACTTGGGC CCTGAAGAAG
     1141 GGCCCAAGTG GCACCGAGTC GGTGCTTTTT TTGAAGCTTG GGCCCGAACA AAAACTCatc
     1201 tcagaagagg atctgaatag cgccgtcgac catcatcatc atcatcattg agtttaaacg
     1261 gtctccagct tggctgtttt ggcggatgag agaagatttt cagcctgata cagattaaat
     1321 cagaacgcag aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc
     1381 cacctgaccc catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt
     1441 ctccccatgc gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa
     1501 gactGGGCCT TTCGTTTTAT CTGTTGTTTG TCGGTGAACT GGATCTTACT TGACAGCTAG
     1561 CTCAGTCCTA GGTATAATAC TAGTAGGACA CCGCAGGATA CCTGGTTTTA GAGCTAGAAA
     1621 TAGCAAGTTA AAATAAGGCT AGTCCGTTAT CAACTTGAAA AAGTGGCACA TGAGGATCAC
     1681 CCATGTGCTT TTTTTGAAGC TTGGGCCCGA ACAAAAACTC atctcagaag aggatctgaa
     1741 tagcgccgtc gaccatcatc atcatcatca ttgagtttaa acggtctcca gcttggctgt
     1801 tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg cagaagcggt
     1861 ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga ccccatgccg
     1921 aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca tgcgagagta
     1981 gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactGGG CCTTTCGTTT
     2041 TATCTGTTGT TTGTCGGTGA ACTGGATCCT TACTCGAGTC TAGACTGCAG gcttcctcgc
     2101 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     2161 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     2221 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     2281 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     2341 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     2401 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     2461 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     2521 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     2581 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     2641 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     2701 ctagaaggac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     2761 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     2821 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     2881 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agAGTACTgc
     2941 ttggattctc accaataaaa aacgcccggc ggcaaccgag cgttctgaac aaatccagat
     3001 ggagttctga ggtcattact ggatctatca acaggagtcc aagcgagctc tcgaacccca
     3061 gagtcccgct cagaagaact cgtcaagaag gcgatagaag gcgatgcgct gcgaatcggg
     3121 a
//