LOCUS       sequence-354808-        8528 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ShHELIX containing a nicking I-AniI fusion to ShTnsB. 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 8528)
  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 8528)
  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_sgRNA_entry (CJT32)
FEATURES             Location/Qualifiers
     source          1..8528
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(95..163,164..955)
                     /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(313..332)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1126..1714
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1615..1634
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(1814..1819,1820..1837,1838..1844)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     RBS             1845..1856
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      8090..8161
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        8175..8209
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     terminator      8464..8491
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     promoter        join(8518..8528,1..94)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
       61 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
      121 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
      181 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
      241 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
      301 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
      361 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
      421 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
      481 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
      541 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
      601 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
      661 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
      721 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
      781 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
      841 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
      901 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
      961 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     1021 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     1081 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     1141 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     1201 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     1261 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     1321 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     1381 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     1441 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     1501 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     1561 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     1621 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     1681 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     1741 gttcctggcc ttttgctggc cttttgctca acgccaagct ccaaaacgat ctcaagaaga
     1801 tcatcttatt aattttacac tttatgcttc cggctcgtat gttgaaagag gagaaaggat
     1861 ctatgggcag cgatttaaca tacgcatatc tagtaggcct gtttgagggt gatggttatt
     1921 tttccattac gaaaaaaggt aaatacctta cctatgagct ggggatcgag ctgagcatca
     1981 aggacgtgca gctgatctat aaaatcaaga agattctggg catcggcatt gtaagcttcc
     2041 gtaaacgcaa tgaaattgag atggtggcgc tgcgcattcg cgataagaac cacctgaaaa
     2101 gcaaaatttt gccaatcttc gagaaatacc cgatgttttc taacaaacaa tacgactatc
     2161 tgcgtttccg caacgctttg ctgagcggta ttatctccct ggaagatctg ccggattaca
     2221 cgcgttctga tgaaccgctg aacagcattg agagcatcat taacaccagc tatttcagtg
     2281 cgtggctggt tggttttatc gaggctgaag gctgcttttc cgtgtacaag ttaaacaagg
     2341 atgacgacta cttgatcgcg agcttcgaca ttgcgcaacg tgacggcgac atcctcatct
     2401 cggccattcg taagtacttg tcattcacca ctaaagttta cttagataaa accaattgta
     2461 gcaagttgaa ggtcacctct gttcgttccg tggaaaacat tatcaaattc ctgcaaaacg
     2521 caccggttaa attgttgggt aatatgaaac tgcagtataa gctgtggctg aagcagctga
     2581 gaaagatttc tcgttacagc gaaaagatca agatcccgag caattatagc ggcagcgaga
     2641 ctcccgggac ctcagagtcc gccacacccg aaagtgggag caacagtcag caaaatcctg
     2701 atttagctgt tcatcccttg gcaattccta tggaaggctt actaggagaa agtgctacaa
     2761 ctcttgagaa gaatgtaatt gccacacaac tctcagagga agcccaagta aagctagagg
     2821 taatccaaag tttactggaa ccctgcgatc gcacaactta tgggcaaaag ttgcgggaag
     2881 cagcagagaa actaaatgta tcgttgcgaa cggtacaaag gttggtgaaa aactgggaac
     2941 aagatggctt agtcggactc actcaaacaa gtagggctga taaaggaaaa caccgcattg
     3001 gtgagttttg ggaaaacttc attaccaaaa cctacaagga gggtaacaag ggaagtaaac
     3061 gtatgacccc taaacaagtt gctctcagag tcgaggctaa agcccgtgaa ttaaaagact
     3121 ctaagccgcc caattacaaa accgtgttac gggtattagc acccattttg gaaaagcaac
     3181 aaaaagccaa gagtatccgc agtcctggtt ggagaggaac tacgctttcg gttaaaaccc
     3241 gtgaaggaaa agatttatcg gttgattaca gtaaccatgt ttggcaatgt gaccataccc
     3301 gcgtggatgt gttgctggta gatcaacatg gtgaaatttt aagtcgtccc tggctaacaa
     3361 cagtaattga tacttactct cgttgcatta tgggtatcaa cttgggcttt gatgcaccca
     3421 gttctggggt agtagcatta gcgttacgcc atgcaattct accaaagcgt tacggttccg
     3481 agtacaaact gcattgtgag tggggaacct atggaaaacc agaacatttt tatactgatg
     3541 gcggtaaaga ctttcgctct aaccacttga gtcagattgg ggcgcaattg ggatttgtct
     3601 gtcatttacg cgatcgccct tctgaaggtg gagtagtaga acgtcccttc aaaacattaa
     3661 atgaccaact attttcaacg cttcctgggt acaccggatc taatgtgcag gaacgcccag
     3721 aagatgcaga gaaggacgca agacttactt tgcgagaact agaacagtta cttgtgcgtt
     3781 acatcgtaga tcgttacaac caaagtattg atgcgcggat gggcgaccaa acgcgctttg
     3841 agcgttggga agcaggattg cctacagtgc cagtaccaat accagaacga gatttggata
     3901 tttgtttaat gaagcagtca cggcgcactg tgcaaagagg tggttgtttg cagtttcaga
     3961 atttaatgta tcggggggaa tatttggcag gttatgccgg agaaactgtc aacttaaggt
     4021 ttgaccccag agacattaca acaattttgg tttatcgcca ggaaaacaat caggaagtat
     4081 ttctgactcg cgctcacgct caaggtttgg agacagagca actggcatta gatgaggctg
     4141 aggcagcaag tcgcagactc cgtaccgcag ggaaaactat cagtaaccaa tcattattgc
     4201 aagaagttgt tgaccgcgat gctcttgtcg ctaccaagaa aagccgtaag gagcgtcaaa
     4261 aattggaaca gactgttttg cgatctgctg ctgttgatga aagtaataga gaatccttgc
     4321 cttctcaaat agttgaacca gatgaagtgg aatctacaga aacggttcac tctcaatacg
     4381 aagacattga ggtgtgggac tatgaacaac ttcgtgaaga atatgggttt taaacaatga
     4441 cagaagctca ggcgatcgcc aagcagttgg gtggggtaaa accggatgat gagtggttac
     4501 aagctgaaat tgctcgtctc aagggtaaga gcattgtgcc tttacagcag gtaaaaactc
     4561 tccatgattg gttagatggc aagcgcaagg caagaaaatc ttgccgagta gttggggaat
     4621 cgagaactgg caagacagtt gcttgtgatg cctacagata caggcacaaa cctcagcagg
     4681 aagctggacg acctccaact gtgcctgtcg tttatattcg acctcaccaa aaatgtggcc
     4741 ccaaggattt gtttaaaaag attactgagt acctcaagta tcgggtaaca aaagggactg
     4801 tatctgattt tcgagatagg acgatagaag tactcaaggg ttgtggcgta gagatgctaa
     4861 ttattgatga agctgaccgt ctcaagcctg aaacttttgc tgatgtgcga gatattgccg
     4921 aagatttagg aattgctgtg gtactggtag gaacagaccg tttggatgcg gtaattaagc
     4981 gggatgagca ggttctcgaa cgctttcggg cgcatcttcg ctttggtaaa ttgtcgggag
     5041 aggattttaa gaacaccgta gaaatgtggg aacaaatggt tttgaaactg ccagtatctt
     5101 ctaatctaaa gagcaaggag atgctacgga ttctcacgtc agcaactgaa ggctacattg
     5161 gtcgccttga tgagattctt agggaagctg caattcgttc cttatcaaga ggattgaaga
     5221 agattgacaa ggctgtttta caggaagtag ctaaggagta caaatgatag aagcaccaga
     5281 tgttaaacct tggctattct tgattaaacc ctatgaaggg gaaagcctga gccactttct
     5341 tggcaggttc agacgtgcca accatttatc cgcaagtgga ttgggtactt tggcaggaat
     5401 tggtgctata gtggcacgtt gggaaagatt tcattttaat cctcgcccta gtcagcaaga
     5461 attggaagcg atcgcatctg tagtagaagt ggatgctcaa aggttagccc agatgttacc
     5521 gcctgctgga gtgggaatgc agcatgagcc aattcgcttg tgtggggctt gttatgccga
     5581 gtcgccttgt caccgaattg aatggcagta caagtcggtg tggaagtgcg atcgccatca
     5641 actcaagatt ttagcaaagt gtccaaactg tcaagcacct tttaaaatgc ctgcgctgtg
     5701 ggaggatggg tgctgtcaca gatgtaggat gccgtttgca gaaatggcaa agctacagaa
     5761 ggtttgatga taaaaccaga aaaaggtgtg aaattaacta agtccctgaa ttgatctggt
     5821 tgtccaaaaa atttgtgcga tcgcatggca agattattcc tactattgat gtggcgtgag
     5881 tcggataact tgctctaatg ggatcccggt taataccaat agatcagata tgagtaataa
     5941 acattacgac ctttacctgt ggtattcact gttggcacga caacaccttt atcgcgccaa
     6001 tactgtagct gacgacgcga acaattagtg atttgagccg cttctgtgct tgtgtagaat
     6061 tttccttcca tagaaatcat cctacagaag tagtttttta aatttacatt ttttaaaagc
     6121 acattatgta ttcatgtgat attatttttt tatgagtcaa ataactattc aagctcgact
     6181 tatttccttt gaatcaaacc gccaacaact ctggaagttg atggcagatt taaacacgcc
     6241 gttaattaac gaactgcttt gccagttagg tcaacacccc gacttcgaga agtggcaaca
     6301 aaagggtaaa ctcccgtcta ccgttgtgag ccagttatgt caacctctca aaactgaccc
     6361 tcgctttgca ggtcagccca gccgtttata tatgtcggca attcatattg tggactacat
     6421 ctacaagtcc tggctggcta tacagaaacg gcttcaacag cagctagatg gaaagacgcg
     6481 ctggctagaa atgctcaata gcgatgctga attagtagaa cttagtggtg acactttaga
     6541 ggctattcgt gtcaaagctg ctgaaatttt ggcaatagct atgccagcat ctgagtcaga
     6601 tagcgcttca cctaaaggga aaaaaggtaa aaaggagaaa aaaccctcat cttctagccc
     6661 taagcgtagt ttatccaaga cattatttga cgcttaccaa gaaacggaag atatcaagag
     6721 ccgtagcgcc atcagctacc tgttaaaaaa tggctgcaaa cttactgaca aagaagaaga
     6781 ttcagaaaaa tttgctaaac gtcgtcgtca agttgaaatc caaattcaaa ggcttaccga
     6841 aaagttaata agtcggatgc ctaaaggtcg agatttgacc aatgctaaat ggttggagac
     6901 actcttgact gctacaacca ctgttgctga agacaacgcc caagccaaac gctggcagga
     6961 tattctgtta actcgatcaa gttctctccc attccccctt gtttttgaaa ccaacgagga
     7021 tatggtttgg tcaaagaatc aaaagggtag gctgtgtgtt cacttcaatg gcttaagcga
     7081 tttaattttt gaggtgtact gcggcaatcg tcaacttcac tggtttcaac gcttcctaga
     7141 agaccaacag actaaacgca aaagcaaaaa tcagcattct agcggcttgt tcacactcag
     7201 aaatggtcat ctagtttggc ttgaaggtga gggtaaaggg gaaccttgga atcttcacca
     7261 cttgaccctt tactgctgtg ttgacaatcg cttgtggaca gaggagggaa cagaaatcgt
     7321 tcgccaagag aaagcagatg aaattactaa attcatcaca aacatgaaga agaaaagcga
     7381 tctaagcgat acacagcaag ctttgattca acgtaaacaa tcaacactta ctcgaataaa
     7441 caattccttt gagcgtccta gccaacccct ttatcaaggt caatcacaca ttttggttgg
     7501 agtaagcctg ggactagaaa aacctgccac agtagcagta gtagatgcga tcgccaacaa
     7561 agtcttggct taccggagta ttaaacaatt acttggcgac aattacgaac tgctaaatcg
     7621 ccagagacga caacagcagt acctatctca cgaacgccac aaagcacaaa aaaacttctc
     7681 tcccaatcaa tttggagcat ctgagttagg gcaacatata gacagattat tagctaaagc
     7741 aattgtagcg ttagcgagaa cctacaaagc tggcagtatt gtcttgccca agttagggga
     7801 tatgcgggag gttgtccaaa gtgaaattca agctatagca gaacaaaaat ttcccggtta
     7861 tattgaaggt cagcaaaaat atgccaaaca gtaccgggtt aatgttcatc ggtggagcta
     7921 cggcagatta attcaaagca ttcaaagtaa agcagctcaa acaggaattg tgattgagga
     7981 gggaaaacaa cctattcgag gtagtcccca cgacaaagca aaggaattag cactttctgc
     8041 ttacaatctc cgcctaacta ggcgaagtta agtaaggatc tccaggcatc aaataaaacg
     8101 aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct
     8161 ctactagaga gaccctgaca gctagctcag tcctaggtat aatgctagca tattaatagc
     8221 gccgcaattc atgctgcttg cagcctctga attttgttaa atgagggtta gtttgactgt
     8281 ataaatacag tcttgctttc tgaccctggt agctgctcac cctgatgctg ctgtcaatag
     8341 acaggatagg tgcgctccca gcaataaggg cgcggatgta ctgctgtagt ggctactgaa
     8401 tcacccccga tcaaggggga accctaaatg ggttgaaagg aagagcgaga tctcgagctc
     8461 ttcggctcac cttcgggtgg gcctttctgc gcaggtggca cttttcgggg aaatgtgcgc
     8521 ggaacccc
//
