LOCUS       sequence-410938-        5450 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of ISCytsp1 TnpB and its reRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5450)
  AUTHORS   Xiang G, Li Y, Sun J, Huo Y, Cao S, Cao Y, Guo Y, Yang L, Cai Y, 
            Zhang YE, Wang H
  TITLE     Evolutionary mining and functional characterization of TnpB 
            nucleases identify efficient miniature genome editors.
  JOURNAL   Nat Biotechnol. 2023 Jun 29. doi: 10.1038/s41587-023-01857-x.
  PUBMED    37386294
REFERENCE   2  (bases 1 to 5450)
  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 Jun 29. doi: 10.1038/s41587-023-01857-x."
COMMENT     Sequence Label: pBad33_ISCytsp1_reRNA
FEATURES             Location/Qualifiers
     source          1..5450
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        24..58
                     /label=BBa_J23108
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.51"
     RBS             74..85
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(1501..1518)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1671..1757
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1849..1876
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1895..1986
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      2444..2899
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2531..2550)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2741..2762
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3455..4114)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     4029..4048
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(4115..4217)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(4609..4626)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4743..5288)
                     /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(4823..4842)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5364..5383
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 atgtttgaca gcttatcatc gatctgacag ctagctcagt cctaggtata atgctagccc
       61 cgggaaaaga tctaaagagg agaaaatgac taaacaaaat aaagcgtaca agttcaggtt
      121 gtatccaaca gatgaacaag cttttattat acgaaaaacc tttgggtgtg ttcgcttcgt
      181 ctacaataaa atgttagctg aacgaataga aacgtatgaa agcctaaaag aagataaaga
      241 agcattgaaa aagataaagc acccaactcc tgctaagtat aaaaaagagt atgaatggct
      301 aaaagaagta gactctttag cattagcgaa cgcacaacta aacttagata aggcatataa
      361 agccttcttc aaagggaatg tcaagtttcc aacgttcaaa aggaaacgac acaaacaaag
      421 ctacacaaca aatgttgtca atggaaatat tcagttgttg gatggtcata tcaaattacc
      481 taagttgaaa ttggtcaaaa ttaaacaaca tcgaaagata ccttcagaat acaaaatcaa
      541 atcctgcaca ttatctatga ccgcatcagg aaaatattat atttctattt tggcagagta
      601 cgaggtggaa attgaaagga aggaagttaa aaaagtcgtt gggttagatt ttgcgatgga
      661 tggattgttt gtggatagcg agggtgagaa agccaattac cctaagtttt atcgacaaaa
      721 acttgaaaaa ttagcaaaag aacaacgtaa actgtctcgt aagaagaaag gttcttccaa
      781 ttggaataaa caacgtattc gagtagctaa cattcaagag aaagtcgcca atcaacggaa
      841 gaactttctt caccatcaat caaaaaaatt agtagcgaac tatgatgctg ttgtgattga
      901 agatttagat atgaaaggaa tgtcacaagc acttaaattc ggtaaaagtg tagctgataa
      961 tggatgggga atgttcactt ctttcttaca gtacaaacta aaagaacaag ggaaacaact
     1021 gatcaaaata gacaaatggt tcccctctac ccaaacctgt tcgcattgtg gaaataccca
     1081 actaatgcca atgaacatca gaatatatgt ttgttcgtgc ggactaaacc tcgatagaga
     1141 ttacaattca gcactcaata tcaaaaaaga aggcattcgc ttattagcaa gtgcctgaga
     1201 gatctttgac agctagctca gtcctaggta taatagatag agattacaat tcagcactca
     1261 atatcaaaaa agaaggcatt cgcttattag caagtgccta aagaaataag agatataaac
     1321 tcttggaaca agagggttag cttggtcaat ttcgtcagct accaaaagtg gcgattaccc
     1381 aagaagcccc cacttcaagc aacccgcaag ggtggtaagt ggtgggagta gttcacgctc
     1441 ggagatcatc attgcgtttt ttaagcttgg ctgttttggc ggatgagaga agattttcag
     1501 cctgatacag attaaatcag aacgcagaag cggtctgata aaacagaatt tgcctggcgg
     1561 cagtagcgcg gtggtcccac ctgaccccat gccgaactca gaagtgaaac gccgtagcgc
     1621 cgatggtagt gtggggtctc cccatgcgag agtagggaac tgccaggcat caaataaaac
     1681 gaaaggctca gtcgaaagac tgggcctttc gttttatctg ttgtttgtcg gtgaacgctc
     1741 tcctgagtag gacaaatccg ccgggagcgg atttgaacgt tgcgaagcaa cggcccggag
     1801 ggtggcgggc aggacgcccg ccataaactg ccaggcatca aattaagcag aaggccatcc
     1861 tgacggatgg cctttttgcg tttctacaaa ctcttttgtt tatttttcta aatacattca
     1921 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     1981 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     2041 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     2101 ggtgcagcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2161 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2221 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2281 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2341 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2401 aactgtcaga ccaagtttac tcatatatac tttagattga tttacgcgcc ctgtagcggc
     2461 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2521 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2581 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2641 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2701 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2761 ggaacaacac tcaaccctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     2821 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     2881 atattaacgt ttacaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     2941 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3001 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3061 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3121 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3181 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3241 agtcaggcat ttgagaagca cacggtcaca ctgcttccgg tagtcaataa accggtaaac
     3301 cagcaataga cataagcggc tatttaacga ccctgccctg aaccgacgac cgggtcgaat
     3361 ttgctttcga atttctgcca ttcatccgct tattatcact tattcaggcg tagcaaccag
     3421 gcgtttaagg gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc
     3481 agtactgttg taattcatta agcattctgc cgacatggaa gccatcacaa acggcatgat
     3541 gaacctgaat cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg
     3601 tgaaaacggg ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac
     3661 tcacccaggg attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg
     3721 ccaggttttc accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat
     3781 cgtcgtggta ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt
     3841 aacaagggtg aacactatcc catatcacca gctcaccgtc tttcattgcc atacggaact
     3901 ccggatgagc attcatcagg cgggcaagaa tgtgaataaa ggccggataa aacttgtgct
     3961 tatttttctt tacggtcttt aaaaaggccg taatatccag ctgaacggtc tggttatagg
     4021 tacattgagc aactgactga aatgcctcaa aatgttcttt acgatgccat tgggatatat
     4081 caacggtggt atatccagtg atttttttct ccattttagc ttccttagct cctgaaaatc
     4141 tcgataactc aaaaaatacg cccggtagtg atcttatttc attatggtga aagttggaac
     4201 ctcttacgtg ccgatcaacg tctcattttc gccaaaagtt ggcccagggc ttcccggtat
     4261 caacagggac accaggattt atttattctg cgaagtgatc ttccgtcaca ggtatttatt
     4321 cggcgcaaag tgcgtcgggt gatgctgcca acttactgat ttagtgtatg atggtgtttt
     4381 tgaggtgctc cagtggcttc tgtttctatc agctgtccct cctgttcagc tactgacggg
     4441 gtggtgcgta acggcaaaag caccgccgga catcagcgct agcggagtgt atactggctt
     4501 actatgttgg cactgatgag ggtgtcagtg aagtgcttca tgtggcagga gaaaaaaggc
     4561 tgcaccggtg cgtcagcaga atatgtgata caggatatat tccgcttcct cgctcactga
     4621 ctcgctacgc tcggtcgttc gactgcggcg agcggaaatg gcttacgaac ggggcggaga
     4681 tttcctggaa gatgccagga agatacttaa cagggaagtg agagggccgc ggcaaagccg
     4741 tttttccata ggctccgccc ccctgacaag catcacgaaa tctgacgctc aaatcagtgg
     4801 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggcgg ctccctcgtg
     4861 cgctctcctg ttcctgcctt tcggtttacc ggtgtcattc cgctgttatg gccgcgtttg
     4921 tctcattcca cgcctgacac tcagttccgg gtaggcagtt cgctccaagc tggactgtat
     4981 gcacgaaccc cccgttcagt ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     5041 caacccggaa agacatgcaa aagcaccact ggcagcagcc actggtaatt gatttagagg
     5101 agttagtctt gaagtcatgc gccggttaag gctaaactga aaggacaagt tttggtgact
     5161 gcgctcctcc aagccagtta cctcggttca aagagttggt agctcagaga accttcgaaa
     5221 aaccgccctg caaggcggtt ttttcgtttt cagagcaaga gattacgcgc agaccaaaac
     5281 gatctcaaga agatcatctt attaatcaga taaaatattt ctaggctcat gagcccgaag
     5341 tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct
     5401 gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggatctgctc
//
