LOCUS       sequence-255015-        3740 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses CRISPR RNA from a T7 promoter for VchCAST system. The 
            CRISPR array encodes a guide RNA with spacer 3 from the native 
            CRISPR..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3740)
  AUTHORS   Klompe SE, Vo PLH, Halpin-Healy TS, Sternberg SH
  TITLE     Transposon-encoded CRISPR-Cas systems direct RNA-guided DNA 
            integration.
  JOURNAL   Nature. 2019 Jul;571(7764):219-225. doi: 10.1038/s41586-019-1323-z. 
            Epub 2019 Jun 12.
  PUBMED    31189177
REFERENCE   2  (bases 1 to 3740)
  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"; doi: 
            "10.1038/s41586-019-1323-z"; journalName: "Nature"; date: "2019-07";
            volume: "571"; issue: "7764"; pages: "219-225"
COMMENT     Sequence Label: pSL0426 (CRISPR with spc3)
FEATURES             Location/Qualifiers
     source          1..3740
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(225..1307)
                     /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     1269..1288
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(1308..1385)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1431..1450
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1431..1449
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1450..1474
                     /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)."
     primer_bind     complement(1625..1643)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1639..1686
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1907..2566)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2481..2500
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2567..2669)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(3061..3078)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3195..3740)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(3275..3294)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca
       61 aatgtagcac ctgaagtcag ccccatacga tataagttgt aattctcatg ttagtcatgc
      121 cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc ggtcgagatc
      181 ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg cccgctttcc
      241 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
      301 gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg caacagctga
      361 ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct ggtttgcccc
      421 agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga gctgtcttcg
      481 gtatcgtcgt atcccactac cgagatgtcc gcaccaacgc gcagcccgga ctcggtaatg
      541 gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt gggaacgatg
      601 ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca gtcgccttcc
      661 cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc agccagacgc
      721 agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg gtgacccaat
      781 gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat aatactgttg
      841 atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca ggcagcttcc
      901 acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact gacgcgttgc
      961 gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc taccatcgac
     1021 accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac aatttgcgac
     1081 ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg tttgcccgcc
     1141 agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc ttccactttt
     1201 tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac ggtctgataa
     1261 gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt caccaccctg
     1321 aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg ccattcgatg
     1381 gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaaat taatacgact
     1441 cactataggg gaattgtgag cggataacaa ttcccctgta gaaataattt tgtttaactt
     1501 taataaggag atataccatg ggtgaactgc cgagtaggta gctgataact tacaggacgc
     1561 tttggcttca ttgcttttca ggtgaactgc cgagtaggta gctgataacc ctaggctgct
     1621 gccaccgctg agcaataact agcataaccc cttggggcct ctaaacgggt cttgaggggt
     1681 tttttgctga aacctcaggc atttgagaag cacacggtca cactgcttcc ggtagtcaat
     1741 aaaccggtaa accagcaata gacataagcg gctatttaac gaccctgccc tgaaccgacg
     1801 accgggtcga atttgctttc gaatttctgc cattcatccg cttattatca cttattcagg
     1861 cgtagcaacc aggcgtttaa gggcaccaat aactgcctta aaaaaattac gccccgccct
     1921 gccactcatc gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac
     1981 aaacggcatg atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat
     2041 atttgcccat agtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa
     2101 aactggtgaa actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt
     2161 tagggaaata ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa
     2221 actgccggaa atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat
     2281 ggaaaacggt gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg
     2341 ccatacggaa ctccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat
     2401 aaaacttgtg cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg
     2461 tctggttata ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc
     2521 attgggatat atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag
     2581 ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt
     2641 gaaagttgga acctcttacg tgccgatcaa cgtctcattt tcgccaaaag ttggcccagg
     2701 gcttcccggt atcaacaggg acaccaggat ttatttattc tgcgaagtga tcttccgtca
     2761 caggtattta ttcggcgcaa agtgcgtcgg gtgatgctgc caacttactg atttagtgta
     2821 tgatggtgtt tttgaggtgc tccagtggct tctgtttcta tcagctgtcc ctcctgttca
     2881 gctactgacg gggtggtgcg taacggcaaa agcaccgccg gacatcagcg ctagcggagt
     2941 gtatactggc ttactatgtt ggcactgatg agggtgtcag tgaagtgctt catgtggcag
     3001 gagaaaaaag gctgcaccgg tgcgtcagca gaatatgtga tacaggatat attccgcttc
     3061 ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa tggcttacga
     3121 acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag tgagagggcc
     3181 gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga aatctgacgc
     3241 tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctggc
     3301 ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat tccgctgtta
     3361 tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag ttcgctccaa
     3421 gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat ccggtaacta
     3481 tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag ccactggtaa
     3541 ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact gaaaggacaa
     3601 gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga
     3661 gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc
     3721 gcagaccaaa acgatctcaa
//
