LOCUS       sequence-358226-        8580 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 3-component ShHELIX containing a Cas12k-TnsC fusion. New 
            sgRNA spacer sequences can be added using Golden Gate assembly (via 
            SapI sites)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8580)
  AUTHORS   Tou CJ, Orr B, Kleinstiver BP
  TITLE     Precise cut-and-paste DNA insertion using engineered type V-K 
            CRISPR-associated transposases.
  JOURNAL   Nat Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01574-x.
  PUBMED    36593413
REFERENCE   2  (bases 1 to 8580)
  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: "Nat 
            Biotechnol. 2023 Jan 2. doi: 10.1038/s41587-022-01574-x."
COMMENT     Sequence Label: pShHELIX(Cas12k-TnsC)-sgRNA_entry (CJT113)
FEATURES             Location/Qualifiers
     source          1..8580
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(103..108,109..126,127..133)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             134..145
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      6431..6502
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        6516..6550
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     terminator      6805..6832
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        6859..6963
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6964..7032,7033..7824)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(7182..7201)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(7995..8580,1..3)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8484..8503
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcaa
       61 cgccaagctc caaaacgatc tcaagaagat catcttatta attttacact ttatgcttcc
      121 ggctcgtatg ttgaaagagg agaaaggatc tatgggcagc gatttaacat acgcatatct
      181 agtaggcctg tttgagggtg atggttattt ttccattacg aaaaaaggta aataccttac
      241 ctatgagctg gggatcgagc tgagcatcaa ggacgtgcag ctgatctata aaatcaagaa
      301 gattctgggc atcggcattg taagcttccg taaacgcaat gaaattgaga tggtggcgct
      361 gcgcattcgc gataagaacc acctgaaaag caaaattttg ccaatcttcg agaaataccc
      421 gatgttttct aacaaacaat acgactatct gcgtttccgc aacgctttgc tgagcggtat
      481 tatctccctg gaagatctgc cggattacac gcgttctgat gaaccgctga acagcattga
      541 gagcatcatt aacaccagct atttcagtgc gtggctggtt ggttttatcg aggctgaagg
      601 ctgcttttcc gtgtacaagt taaacaagga tgacgactac ttgatcgcga gcttcgacat
      661 tgcgcaacgt gacggcgaca tcctcatctc ggccattcgt aagtacttgt cattcaccac
      721 taaagtttac ttagataaaa ccaattgtag caagttgaag gtcacctctg ttcgttccgt
      781 ggaaaacatt atcaaattcc tgcaaaacgc accggttaaa ttgttgggta atatgaaact
      841 gcagtataag ctgtggctga agcagctgag aaagatttct cgttacagcg aaaagatcaa
      901 gatcccgagc aattatagcg gcagcgagac tcccgggacc tcagagtccg ccacacccga
      961 aagtgggagc aacagtcagc aaaatcctga tttagctgtt catcccttgg caattcctat
     1021 ggaaggctta ctaggagaaa gtgctacaac tcttgagaag aatgtaattg ccacacaact
     1081 ctcagaggaa gcccaagtaa agctagaggt aatccaaagt ttactggaac cctgcgatcg
     1141 cacaacttat gggcaaaagt tgcgggaagc agcagagaaa ctaaatgtat cgttgcgaac
     1201 ggtacaaagg ttggtgaaaa actgggaaca agatggctta gtcggactca ctcaaacaag
     1261 tagggctgat aaaggaaaac accgcattgg tgagttttgg gaaaacttca ttaccaaaac
     1321 ctacaaggag ggtaacaagg gaagtaaacg tatgacccct aaacaagttg ctctcagagt
     1381 cgaggctaaa gcccgtgaat taaaagactc taagccgccc aattacaaaa ccgtgttacg
     1441 ggtattagca cccattttgg aaaagcaaca aaaagccaag agtatccgca gtcctggttg
     1501 gagaggaact acgctttcgg ttaaaacccg tgaaggaaaa gatttatcgg ttgattacag
     1561 taaccatgtt tggcaatgtg accatacccg cgtggatgtg ttgctggtag atcaacatgg
     1621 tgaaatttta agtcgtccct ggctaacaac agtaattgat acttactctc gttgcattat
     1681 gggtatcaac ttgggctttg atgcacccag ttctggggta gtagcattag cgttacgcca
     1741 tgcaattcta ccaaagcgtt acggttccga gtacaaactg cattgtgagt ggggaaccta
     1801 tggaaaacca gaacattttt atactgatgg cggtaaagac tttcgctcta accacttgag
     1861 tcagattggg gcgcaattgg gatttgtctg tcatttacgc gatcgccctt ctgaaggtgg
     1921 agtagtagaa cgtcccttca aaacattaaa tgaccaacta ttttcaacgc ttcctgggta
     1981 caccggatct aatgtgcagg aacgcccaga agatgcagag aaggacgcaa gacttacttt
     2041 gcgagaacta gaacagttac ttgtgcgtta catcgtagat cgttacaacc aaagtattga
     2101 tgcgcggatg ggcgaccaaa cgcgctttga gcgttgggaa gcaggattgc ctacagtgcc
     2161 agtaccaata ccagaacgag atttggatat ttgtttaatg aagcagtcac ggcgcactgt
     2221 gcaaagaggt ggttgtttgc agtttcagaa tttaatgtat cggggggaat atttggcagg
     2281 ttatgccgga gaaactgtca acttaaggtt tgaccccaga gacattacaa caattttggt
     2341 ttatcgccag gaaaacaatc aggaagtatt tctgactcgc gctcacgctc aaggtttgga
     2401 gacagagcaa ctggcattag atgaggctga ggcagcaagt cgcagactcc gtaccgcagg
     2461 gaaaactatc agtaaccaat cattattgca agaagttgtt gaccgcgatg ctcttgtcgc
     2521 taccaagaaa agccgtaagg agcgtcaaaa attggaacag actgttttgc gatctgctgc
     2581 tgttgatgaa agtaatagag aatccttgcc ttctcaaata gttgaaccag atgaagtgga
     2641 atctacagaa acggttcact ctcaatacga agacattgag gtgtgggact atgaacaact
     2701 tcgtgaagaa tatgggtttt aaacaatgat agaagcacca gatgttaaac cttggctatt
     2761 cttgattaaa ccctatgaag gggaaagcct gagccacttt cttggcaggt tcagacgtgc
     2821 caaccattta tccgcaagtg gattgggtac tttggcagga attggtgcta tagtggcacg
     2881 ttgggaaaga tttcatttta atcctcgccc tagtcagcaa gaattggaag cgatcgcatc
     2941 tgtagtagaa gtggatgctc aaaggttagc ccagatgtta ccgcctgctg gagtgggaat
     3001 gcagcatgag ccaattcgct tgtgtggggc ttgttatgcc gagtcgcctt gtcaccgaat
     3061 tgaatggcag tacaagtcgg tgtggaagtg cgatcgccat caactcaaga ttttagcaaa
     3121 gtgtccaaac tgtcaagcac cttttaaaat gcctgcgctg tgggaggatg ggtgctgtca
     3181 cagatgtagg atgccgtttg cagaaatggc aaagctacag aaggtttgat gataaaacca
     3241 gaaaaaggtg tgaaattaac taagtccctg aattgatctg gttgtccaaa aaatttgtgc
     3301 gatcgcatgg caagattatt cctactattg atgtggcgtg agtcggataa cttgctctaa
     3361 tgggatcccg gttaatacca atagatcaga tatgagtaat aaacattacg acctttacct
     3421 gtggtattca ctgttggcac gacaacacct ttatcgcgcc aatactgtag ctgacgacgc
     3481 gaacaattag tgatttgagc cgcttctgtg cttgtgtaga attttccttc catagaaatc
     3541 atcctacaga agtagttttt taaatttaca ttttttaaaa gcacattatg tattcatgtg
     3601 atattatttt tttatgagtc aaataactat tcaagctcga cttatttcct ttgaatcaaa
     3661 ccgccaacaa ctctggaagt tgatggcaga tttaaacacg ccgttaatta acgaactgct
     3721 ttgccagtta ggtcaacacc ccgacttcga gaagtggcaa caaaagggta aactcccgtc
     3781 taccgttgtg agccagttat gtcaacctct caaaactgac cctcgctttg caggtcagcc
     3841 cagccgttta tatatgtcgg caattcatat tgtggactac atctacaagt cctggctggc
     3901 tatacagaaa cggcttcaac agcagctaga tggaaagacg cgctggctag aaatgctcaa
     3961 tagcgatgct gaattagtag aacttagtgg tgacacttta gaggctattc gtgtcaaagc
     4021 tgctgaaatt ttggcaatag ctatgccagc atctgagtca gatagcgctt cacctaaagg
     4081 gaaaaaaggt aaaaaggaga aaaaaccctc atcttctagc cctaagcgta gtttatccaa
     4141 gacattattt gacgcttacc aagaaacgga agatatcaag agccgtagcg ccatcagcta
     4201 cctgttaaaa aatggctgca aacttactga caaagaagaa gattcagaaa aatttgctaa
     4261 acgtcgtcgt caagttgaaa tccaaattca aaggcttacc gaaaagttaa taagtcggat
     4321 gcctaaaggt cgagatttga ccaatgctaa atggttggag acactcttga ctgctacaac
     4381 cactgttgct gaagacaacg cccaagccaa acgctggcag gatattctgt taactcgatc
     4441 aagttctctc ccattccccc ttgtttttga aaccaacgag gatatggttt ggtcaaagaa
     4501 tcaaaagggt aggctgtgtg ttcacttcaa tggcttaagc gatttaattt ttgaggtgta
     4561 ctgcggcaat cgtcaacttc actggtttca acgcttccta gaagaccaac agactaaacg
     4621 caaaagcaaa aatcagcatt ctagcggctt gttcacactc agaaatggtc atctagtttg
     4681 gcttgaaggt gagggtaaag gggaaccttg gaatcttcac cacttgaccc tttactgctg
     4741 tgttgacaat cgcttgtgga cagaggaggg aacagaaatc gttcgccaag agaaagcaga
     4801 tgaaattact aaattcatca caaacatgaa gaagaaaagc gatctaagcg atacacagca
     4861 agctttgatt caacgtaaac aatcaacact tactcgaata aacaattcct ttgagcgtcc
     4921 tagccaaccc ctttatcaag gtcaatcaca cattttggtt ggagtaagcc tgggactaga
     4981 aaaacctgcc acagtagcag tagtagatgc gatcgccaac aaagtcttgg cttaccggag
     5041 tattaaacaa ttacttggcg acaattacga actgctaaat cgccagagac gacaacagca
     5101 gtacctatct cacgaacgcc acaaagcaca aaaaaacttc tctcccaatc aatttggagc
     5161 atctgagtta gggcaacata tagacagatt attagctaaa gcaattgtag cgttagcgag
     5221 aacctacaaa gctggcagta ttgtcttgcc caagttaggg gatatgcggg aggttgtcca
     5281 aagtgaaatt caagctatag cagaacaaaa atttcccggt tatattgaag gtcagcaaaa
     5341 atatgccaaa cagtaccggg ttaatgttca tcggtggagc tacggcagat taattcaaag
     5401 cattcaaagt aaagcagctc aaacaggaat tgtgattgag gagggaaaac aacctattcg
     5461 aggtagtccc cacgacaaag caaaggaatt agcactttct gcttacaatc tccgcctaac
     5521 taggcgaagt agcggcagcg agactcccgg gacctcagag tccgccacac ccgaaagtgg
     5581 gagcacagaa gctcaggcga tcgccaagca gttgggtggg gtaaaaccgg atgatgagtg
     5641 gttacaagct gaaattgctc gtctcaaggg taagagcatt gtgcctttac agcaggtaaa
     5701 aactctccat gattggttag atggcaagcg caaggcaaga aaatcttgcc gagtagttgg
     5761 ggaatcgaga actggcaaga cagttgcttg tgatgcctac agatacaggc acaaacctca
     5821 gcaggaagct ggacgacctc caactgtgcc tgtcgtttat attcgacctc accaaaaatg
     5881 tggccccaag gatttgttta aaaagattac tgagtacctc aagtatcggg taacaaaagg
     5941 gactgtatct gattttcgag ataggacgat agaagtactc aagggttgtg gcgtagagat
     6001 gctaattatt gatgaagctg accgtctcaa gcctgaaact tttgctgatg tgcgagatat
     6061 tgccgaagat ttaggaattg ctgtggtact ggtaggaaca gaccgtttgg atgcggtaat
     6121 taagcgggat gagcaggttc tcgaacgctt tcgggcgcat cttcgctttg gtaaattgtc
     6181 gggagaggat tttaagaaca ccgtagaaat gtgggaacaa atggttttga aactgccagt
     6241 atcttctaat ctaaagagca aggagatgct acggattctc acgtcagcaa ctgaaggcta
     6301 cattggtcgc cttgatgaga ttcttaggga agctgcaatt cgttccttat caagaggatt
     6361 gaagaagatt gacaaggctg ttttacagga agtagctaag gagtacaaat aagtaaggat
     6421 ctccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     6481 ttgtttgtcg gtgaacgctc tctactagag agaccctgac agctagctca gtcctaggta
     6541 taatgctagc atattaatag cgccgcaatt catgctgctt gcagcctctg aattttgtta
     6601 aatgagggtt agtttgactg tataaataca gtcttgcttt ctgaccctgg tagctgctca
     6661 ccctgatgct gctgtcaata gacaggatag gtgcgctccc agcaataagg gcgcggatgt
     6721 actgctgtag tggctactga atcacccccg atcaaggggg aaccctaaat gggttgaaag
     6781 gaagagcgag atctcgagct cttcggctca ccttcgggtg ggcctttctg cgcaggtggc
     6841 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     6901 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     6961 agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt
     7021 cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt
     7081 gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga gagttttcgc
     7141 cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta
     7201 tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac
     7261 ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa
     7321 ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg
     7381 atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc
     7441 cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg
     7501 atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta
     7561 gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg accacttctg
     7621 cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg
     7681 tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc
     7741 tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt
     7801 gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt
     7861 gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc
     7921 atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag
     7981 atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa
     8041 aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg
     8101 aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt gtagccgtag
     8161 ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg
     8221 ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga
     8281 tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc
     8341 ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc
     8401 acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga
     8461 gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt
     8521 cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg
//
