LOCUS       sequence-248518-        8150 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses ShCAST and a crRNA targeting PSP1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8150)
  AUTHORS   Strecker J, Ladha A, Gardner Z, Schmid-Burgk JL, Makarova KS, Koonin
            EV, Zhang F
  TITLE     RNA-guided DNA insertion with CRISPR-associated transposases.
  JOURNAL   Science. 2019 Jul 5;365(6448):48-53. doi: 10.1126/science.aax9181. 
            Epub 2019 Jun 6.
  PUBMED    31171706
REFERENCE   2  (bases 1 to 8150)
  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: 
            "10.1126/science.aax9181"; journalName: "Science"; date: "2019-07-5-
            5"; volume: "365"; issue: "6448"; pages: "48-53"
COMMENT     Sequence Label: pHelper_ShCAST
FEATURES             Location/Qualifiers
     source          1..8150
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             1..12
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        5857..5891
                     /label=J23119 promoter
                     /note="bacterial promoter (Registry of Standard Biological 
                     Parts BBa_J23119)"
     primer_bind     complement(6031..6050)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6150..6172
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6210..6228)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6296..6400
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6401..6469,6470..7261)
                     /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(6619..6638)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7432..8020
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7921..7940
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        join(8120..8125,8126..8143,8144..8150)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
ORIGIN
        1 aaagaggaga aaggatctat gaacagtcag caaaatcctg atttagctgt tcatcccttg
       61 gcaattccta tggaaggctt actaggagaa agtgctacaa ctcttgagaa gaatgtaatt
      121 gccacacaac tctcagagga agcccaagta aagctagagg taatccaaag tttactggaa
      181 ccctgcgatc gcacaactta tgggcaaaag ttgcgggaag cagcagagaa actaaatgta
      241 tcgttgcgaa cggtacaaag gttggtgaaa aactgggaac aagatggctt agtcggactc
      301 actcaaacaa gtagggctga taaaggaaaa caccgcattg gtgagttttg ggaaaacttc
      361 attaccaaaa cctacaagga gggtaacaag ggaagtaaac gtatgacccc taaacaagtt
      421 gctctcagag tcgaggctaa agcccgtgaa ttaaaagact ctaagccgcc caattacaaa
      481 accgtgttac gggtattagc acccattttg gaaaagcaac aaaaagccaa gagtatccgc
      541 agtcctggtt ggagaggaac tacgctttcg gttaaaaccc gtgaaggaaa agatttatcg
      601 gttgattaca gtaaccatgt ttggcaatgt gaccataccc gcgtggatgt gttgctggta
      661 gatcaacatg gtgaaatttt aagtcgtccc tggctaacaa cagtaattga tacttactct
      721 cgttgcatta tgggtatcaa cttgggcttt gatgcaccca gttctggggt agtagcatta
      781 gcgttacgcc atgcaattct accaaagcgt tacggttccg agtacaaact gcattgtgag
      841 tggggaacct atggaaaacc agaacatttt tatactgatg gcggtaaaga ctttcgctct
      901 aaccacttga gtcagattgg ggcgcaattg ggatttgtct gtcatttacg cgatcgccct
      961 tctgaaggtg gagtagtaga acgtcccttc aaaacattaa atgaccaact attttcaacg
     1021 cttcctgggt acaccggatc taatgtgcag gaacgcccag aagatgcaga gaaggacgca
     1081 agacttactt tgcgagaact agaacagtta cttgtgcgtt acatcgtaga tcgttacaac
     1141 caaagtattg atgcgcggat gggcgaccaa acgcgctttg agcgttggga agcaggattg
     1201 cctacagtgc cagtaccaat accagaacga gatttggata tttgtttaat gaagcagtca
     1261 cggcgcactg tgcaaagagg tggttgtttg cagtttcaga atttaatgta tcggggggaa
     1321 tatttggcag gttatgccgg agaaactgtc aacttaaggt ttgaccccag agacattaca
     1381 acaattttgg tttatcgcca ggaaaacaat caggaagtat ttctgactcg cgctcacgct
     1441 caaggtttgg agacagagca actggcatta gatgaggctg aggcagcaag tcgcagactc
     1501 cgtaccgcag ggaaaactat cagtaaccaa tcattattgc aagaagttgt tgaccgcgat
     1561 gctcttgtcg ctaccaagaa aagccgtaag gagcgtcaaa aattggaaca gactgttttg
     1621 cgatctgctg ctgttgatga aagtaataga gaatccttgc cttctcaaat agttgaacca
     1681 gatgaagtgg aatctacaga aacggttcac tctcaatacg aagacattga ggtgtgggac
     1741 tatgaacaac ttcgtgaaga atatgggttt taaacaatga cagaagctca ggcgatcgcc
     1801 aagcagttgg gtggggtaaa accggatgat gagtggttac aagctgaaat tgctcgtctc
     1861 aagggtaaga gcattgtgcc tttacagcag gtaaaaactc tccatgattg gttagatggc
     1921 aagcgcaagg caagaaaatc ttgccgagta gttggggaat cgagaactgg caagacagtt
     1981 gcttgtgatg cctacagata caggcacaaa cctcagcagg aagctggacg acctccaact
     2041 gtgcctgtcg tttatattcg acctcaccaa aaatgtggcc ccaaggattt gtttaaaaag
     2101 attactgagt acctcaagta tcgggtaaca aaagggactg tatctgattt tcgagatagg
     2161 acgatagaag tactcaaggg ttgtggcgta gagatgctaa ttattgatga agctgaccgt
     2221 ctcaagcctg aaacttttgc tgatgtgcga gatattgccg aagatttagg aattgctgtg
     2281 gtactggtag gaacagaccg tttggatgcg gtaattaagc gggatgagca ggttctcgaa
     2341 cgctttcggg cgcatcttcg ctttggtaaa ttgtcgggag aggattttaa gaacaccgta
     2401 gaaatgtggg aacaaatggt tttgaaactg ccagtatctt ctaatctaaa gagcaaggag
     2461 atgctacgga ttctcacgtc agcaactgaa ggctacattg gtcgccttga tgagattctt
     2521 agggaagctg caattcgttc cttatcaaga ggattgaaga agattgacaa ggctgtttta
     2581 caggaagtag ctaaggagta caaatgatag aagcaccaga tgttaaacct tggctattct
     2641 tgattaaacc ctatgaaggg gaaagcctga gccactttct tggcaggttc agacgtgcca
     2701 accatttatc cgcaagtgga ttgggtactt tggcaggaat tggtgctata gtggcacgtt
     2761 gggaaagatt tcattttaat cctcgcccta gtcagcaaga attggaagcg atcgcatctg
     2821 tagtagaagt ggatgctcaa aggttagccc agatgttacc gcctgctgga gtgggaatgc
     2881 agcatgagcc aattcgcttg tgtggggctt gttatgccga gtcgccttgt caccgaattg
     2941 aatggcagta caagtcggtg tggaagtgcg atcgccatca actcaagatt ttagcaaagt
     3001 gtccaaactg tcaagcacct tttaaaatgc ctgcgctgtg ggaggatggg tgctgtcaca
     3061 gatgtaggat gccgtttgca gaaatggcaa agctacagaa ggtttgatga taaaaccaga
     3121 aaaaggtgtg aaattaacta agtccctgaa ttgatctggt tgtccaaaaa atttgtgcga
     3181 tcgcatggca agattattcc tactattgat gtggcgtgag tcggataact tgctctaatg
     3241 ggatcccggt taataccaat agatcagata tgagtaataa acattacgac ctttacctgt
     3301 ggtattcact gttggcacga caacaccttt atcgcgccaa tactgtagct gacgacgcga
     3361 acaattagtg atttgagccg cttctgtgct tgtgtagaat tttccttcca tagaaatcat
     3421 cctacagaag tagtttttta aatttacatt ttttaaaagc acattatgta ttcatgtgat
     3481 attatttttt tatgagtcaa ataactattc aagctcgact tatttccttt gaatcaaacc
     3541 gccaacaact ctggaagttg atggcagatt taaacacgcc gttaattaac gaactgcttt
     3601 gccagttagg tcaacacccc gacttcgaga agtggcaaca aaagggtaaa ctcccgtcta
     3661 ccgttgtgag ccagttatgt caacctctca aaactgaccc tcgctttgca ggtcagccca
     3721 gccgtttata tatgtcggca attcatattg tggactacat ctacaagtcc tggctggcta
     3781 tacagaaacg gcttcaacag cagctagatg gaaagacgcg ctggctagaa atgctcaata
     3841 gcgatgctga attagtagaa cttagtggtg acactttaga ggctattcgt gtcaaagctg
     3901 ctgaaatttt ggcaatagct atgccagcat ctgagtcaga tagcgcttca cctaaaggga
     3961 aaaaaggtaa aaaggagaaa aaaccctcat cttctagccc taagcgtagt ttatccaaga
     4021 cattatttga cgcttaccaa gaaacggaag atatcaagag ccgtagcgcc atcagctacc
     4081 tgttaaaaaa tggctgcaaa cttactgaca aagaagaaga ttcagaaaaa tttgctaaac
     4141 gtcgtcgtca agttgaaatc caaattcaaa ggcttaccga aaagttaata agtcggatgc
     4201 ctaaaggtcg agatttgacc aatgctaaat ggttggagac actcttgact gctacaacca
     4261 ctgttgctga agacaacgcc caagccaaac gctggcagga tattctgtta actcgatcaa
     4321 gttctctccc attccccctt gtttttgaaa ccaacgagga tatggtttgg tcaaagaatc
     4381 aaaagggtag gctgtgtgtt cacttcaatg gcttaagcga tttaattttt gaggtgtact
     4441 gcggcaatcg tcaacttcac tggtttcaac gcttcctaga agaccaacag actaaacgca
     4501 aaagcaaaaa tcagcattct agcggcttgt tcacactcag aaatggtcat ctagtttggc
     4561 ttgaaggtga gggtaaaggg gaaccttgga atcttcacca cttgaccctt tactgctgtg
     4621 ttgacaatcg cttgtggaca gaggagggaa cagaaatcgt tcgccaagag aaagcagatg
     4681 aaattactaa attcatcaca aacatgaaga agaaaagcga tctaagcgat acacagcaag
     4741 ctttgattca acgtaaacaa tcaacactta ctcgaataaa caattccttt gagcgtccta
     4801 gccaacccct ttatcaaggt caatcacaca ttttggttgg agtaagcctg ggactagaaa
     4861 aacctgccac agtagcagta gtagatgcga tcgccaacaa agtcttggct taccggagta
     4921 ttaaacaatt acttggcgac aattacgaac tgctaaatcg ccagagacga caacagcagt
     4981 acctatctca cgaacgccac aaagcacaaa aaaacttctc tcccaatcaa tttggagcat
     5041 ctgagttagg gcaacatata gacagattat tagctaaagc aattgtagcg ttagcgagaa
     5101 cctacaaagc tggcagtatt gtcttgccca agttagggga tatgcgggag gttgtccaaa
     5161 gtgaaattca agctatagca gaacaaaaat ttcccggtta tattgaaggt cagcaaaaat
     5221 atgccaaaca gtaccgggtt aatgttcatc ggtggagcta cggcagatta attcaaagca
     5281 ttcaaagtaa agcagctcaa acaggaattg tgattgagga gggaaaacaa cctattcgag
     5341 gtagtcccca cgacaaagca aaggaattag cactttctgc ttacaatctc cgcctaacta
     5401 ggcgaagtta acaaatatct gaaccttgat aatagaatat taatagcgcc gcaattcatg
     5461 ctgcttgcag cctctgaatt ttgttaaatg agggttagtt tgactgtata aatacagtct
     5521 tgctttctga ccctggtagc tgctcaccct gatgctgctg tcaatagaca ggataggtgc
     5581 gctcccagca ataagggcgc ggatgtactg ctgtagtggc tactgaatca cccccgatca
     5641 agggggaacc ctccccaatt cttcatttga aggactaaaa tcaaggcaaa atttctaaga
     5701 gatccgcgca agttccaaat accttatctc gtcttgattt catctttttt taaccaaggc
     5761 atgattcttg agactgaggc tcaaatgaga aattgggaaa catccgcgct gaagatatct
     5821 ggaaaggttg gctcacaaca ttttgtaact gggtagttga cagctagctc agtcctaggt
     5881 ataatgctag cgtggcaaca accttccagg tactaggtgg gttgaaagga gaagtcattt
     5941 aataaggcca ctgttaaaag tggcaacaac cttccaggta ctaggtgggt tgaaagcggc
     6001 cgacgcgctg ggctactggt gcactctcag tacaatctgc tctgatgccg catagttaag
     6061 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     6121 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     6181 gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     6241 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     6301 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     6361 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     6421 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     6481 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     6541 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     6601 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     6661 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     6721 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     6781 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     6841 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     6901 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     6961 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     7021 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     7081 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     7141 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     7201 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     7261 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     7321 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     7381 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     7441 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     7501 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     7561 gcagatacca aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     7621 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     7681 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     7741 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     7801 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     7861 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     7921 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     7981 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     8041 tttacggttc ctggcctttt gctggccttt tgctcaacgc caagctccaa aacgatctca
     8101 agaagatcat cttattaatt ttacacttta tgcttccggc tcgtatgttg
//
