LOCUS       sequence-182491-        4134 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Spectinomycin resistant plasmid with a CDF origin that expresses 
            Csy4 when IPTG induced, for coexpression with Series 371 vectors.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4134)
  TITLE     Series-371: N- or C-terminal Tag E.coli T7 Expression Vectors with 
            Biobrick Polycistronic Restriction Sites and Csy4 cleavage
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4134)
  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: 
            "Unpublished"
COMMENT     Sequence Label: 13SA-csy4
FEATURES             Location/Qualifiers
     source          1..4134
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(3..1085)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     1047..1066
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1086..1163)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1209..1228
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1209..1227
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1232..1255
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             1308..1330
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1339..1902
                     /codon_start=1
                     /product="CRISPR-associated endoribonuclease responsible 
                     for pre-crRNA processing in Pseudomonas aeruginosa 
                     (Haurwitz et al., 2010)"
                     /label=Csy4
                     /translation="MDHYLDIRLRPDPEFPPAQLMSVLFGKLHQALVAQGGDRIGVSFP
                     DLDESRSRLGERLRIHASADDLRALLARPWLEGLRDHLQFGEPAVVPHPTPYRQVSRVQ
                     AKSNPERLRRRLMRRHDLSEEEARKRIPDTVARALDLPFVTLRSQSTGQHFRLFIRHGP
                     LQVTAEEGGFTCYGLSKGGFVPWF"
     primer_bind     complement(2027..2045)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2041..2088
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(2259..3050)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(3051..3142)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3190..3928)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
ORIGIN
        1 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
       61 aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt
      121 gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg
      181 tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata
      241 taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatgtccgc accaacgcgc
      301 agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc
      361 atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg
      421 gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta
      481 tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg
      541 atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg
      601 gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca
      661 ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc
      721 agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg
      781 cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc
      841 gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc
      901 aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc
      961 atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg
     1021 cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt
     1081 ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag
     1141 gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat
     1201 taggaaatta atacgactca ctatagggag agaattgtga gcggataaca attcccctgt
     1261 agattaatta agcggccgcc ctgcaggact cgagttctag aaataatttt gtttaacttt
     1321 aagaaggaga tatagatcat ggaccactac ctcgacattc gcttgcgacc ggacccggaa
     1381 tttcccccgg cgcaactcat gagcgtgctc ttcggcaagc tccaccaggc cctggtggca
     1441 cagggcgggg acaggatcgg cgtgagcttc cccgacctcg acgaaagccg ctcccggctg
     1501 ggcgagcgcc tgcgcattca tgcctcggcg gacgaccttc gtgccctgct cgcccggccc
     1561 tggctggaag ggttgcggga ccatctgcaa ttcggagaac cggcagtcgt gcctcacccc
     1621 acaccgtacc gtcaggtcag tcgggttcag gcgaaaagca atccggaacg cctgcggcgg
     1681 cggctcatgc gccggcacga tctgagtgag gaggaggctc ggaaacgcat tcccgatacg
     1741 gtcgcgagag ccttggacct gcccttcgtc acgctacgca gccagagcac cggacagcac
     1801 ttccgtctct tcatccgcca cgggccgttg caggtgacgg cagaggaagg aggattcacc
     1861 tgttacgggt tgagcaaagg aggtttcgtt ccctggttct aataagatcc caactccata
     1921 aggatccgcg atcgcggcgc gccacctggt ggccggccgg taccacgcgt gcgcgctgat
     1981 ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa
     2041 ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaacctcag
     2101 gcatttgaga agcacacggt cacactgctt ccggtagtca ataaaccggt aaaccagcaa
     2161 tagacataag cggctattta acgaccctgc cctgaaccga cgaccgggtc atcgtggccg
     2221 gatcttgcgg cccctcggct tgaacgaatt gttagacatt atttgccgac taccttggtg
     2281 atctcgcctt tcacgtaatg gacaaattct tccaactgat ctgcgcgcga ggccaagcga
     2341 tcttcttctt gtccaagata agcctgtcta gcttcaagta tgacgggctg atactgggcc
     2401 ggcaggcgct ccattgccca gtcggcagcg acatccttcg gcgcgatttt gccggttact
     2461 gcgctgtacc aaatgcggga caacgtaagc actacatttc gctcatcgcc agcccagtcg
     2521 ggcggcgagt tccatagcgt taaggtttca tttagcgcct caaatagatc ctgttcagga
     2581 accggatcaa agagttcctc cgccgctgga cctaccaagg caacgctatg ttctcttgct
     2641 tttgtcagca agatagccag atcaatgtcg atcgtggctg gctcgaagat acctgcaaga
     2701 atgtcattgc gctgccattc tccaaattgc agttcgcgct tagctggata acgccacgga
     2761 atgatgtcgt cgtgcacaac aatggtgact tctacagcgc ggagaatctc gctctctcca
     2821 ggggaagccg aagtttccaa aaggtcgttg atcaaagctc gccgcgttgt ttcatcaagc
     2881 cttacggtca ccgtaaccag caaatcaata tcactgtgcg gcttcaggcc gccatccact
     2941 gcggagccgt acaaatgtac ggccagcaac gtcggttcga gatggcgctc gatgacgcca
     3001 actacctctg atagttgagt cgatacttcg gcgatcaccg cttccctcat actcttcctt
     3061 tttcattatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     3121 tgtatttaga aaaataaaca aatagctagc tcactcggtc gctacgctcc gggcgtgaga
     3181 ctgcggcggg cgctgcggac acatacaaag ttacccacag attccgtgga taagcagggg
     3241 actaacatgt gaggcaaaac agcagggccg cgccggtggc gtttttccat aggctccgcc
     3301 ctcctgccag agttcacata aacagacgct tttccggtgc atctgtggga gccgtgaggc
     3361 tcaaccatga atctgacagt acgggcgaaa cccgacagga cttaaagatc cccaccgttt
     3421 ccggcgggtc gctccctctt gcgctctcct gttccgaccc tgccgtttac cggatacctg
     3481 ttccgccttt ctcccttacg ggaagtgtgg cgctttctca tagctcacac actggtatct
     3541 cggctcggtg taggtcgttc gctccaagct gggctgtaag caagaactcc ccgttcagcc
     3601 cgactgctgc gccttatccg gtaactgttc acttgagtcc aacccggaaa agcacggtaa
     3661 aacgccactg gcagcagcca ttggtaactg ggagttcgca gaggatttgt ttagctaaac
     3721 acgcggttgc tcttgaagtg tgcgccaaag tccggctaca ctggaaggac agatttggtt
     3781 gctgtgctct gcgaaagcca gttaccacgg ttaagcagtt ccccaactga cttaaccttc
     3841 gatcaaacca cctccccagg tggttttttc gtttacaggg caaaagatta cgcgcagaaa
     3901 aaaaggatct caagaagatc ctttgatctt ttctactgaa ccgctcttat ttcagtgcaa
     3961 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     4021 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     4081 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgc
//
