LOCUS       sequence-363180-        6328 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSwap plasmid part containing the T1 sgRNA module for the Swap and 
            Drop recombination system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6328)
  AUTHORS   Teufel M, Klein CA, Mager M, Sobetzko P
  TITLE     A multifunctional system for genome editing and large-scale 
            interspecies gene transfer.
  JOURNAL   Nat Commun. 2022 Jun 14;13(1):3430. doi: 10.1038/s41467-022-30843-1.
  PUBMED    35701417
REFERENCE   2  (bases 1 to 6328)
  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 
            Commun."; date: "2022-06-14"; volume: "13(1):3430. doi"; pages: " 
            10.1038/s41467-022-30843-1"
COMMENT     Sequence Label: pSWAP_Entry_T1
FEATURES             Location/Qualifiers
     source          1..6328
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    24..42
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    126..147
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(162..167,168..185,186..192)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    200..216
                     /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     205..227
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     224..240
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     224..240
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    253..309
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(310..327)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(310..326)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(319..341)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(535..554)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             901..1206
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     1085..1104
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     misc_RNA        1314..1389
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     primer_bind     complement(4516..4534)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4602..4706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4707..5501
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
     rep_origin      complement(5561..6149)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5641..5660)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    join(6327..6328,1..17)
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
ORIGIN
        1 cctatcagtg atagagattg acatccctat cagtgataga gatactgagc actgagaccc
       61 agtggtatgg gcagctggca cgacaggttt cccgactgga aagcgggcag tgagcgcaac
      121 gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt tatgcttccg
      181 gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
      241 catgattacg ccaagcttgc atgcctgcag gtcgactcta gaggatcccc gggtaccgag
      301 ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca
      361 acttaatcgc cttgcaggac atcccccttt cgccagctgg cgtaatagcg aagaggcccg
      421 caccgatcgc ccttcccaac agttgcgcag cctgaatggc gaatggcgcc tgatgcggta
      481 ttttctcctt acgcatctgt gcggtatttc acaccgcata tggtgcactc tcagtacaat
      541 ctgctctgat gccgcatagt cgcgtggatc cggcttacta aaagccagat aacagtatgc
      601 gtatttgcgc gctgattttt gcggtataag aatatatact gatatgtata cccgaagtat
      661 gtcaaaaaga ggtatgctat gaagcagcgt attacagtga cagttgacag cgacagctat
      721 cagttgctca aggcatatat gatgtcaata tctccggtct ggtaagcaca accatgcaga
      781 atgaagcccg tcgtctgcgt gccgaacgct ggaaagcgga aaatcaggaa gggatggctg
      841 aggtcgcccg gtttattgaa atgaacggct cttttgctga cgagaacagg ggctggtgaa
      901 atgcagttta aggtttacac ctataaaaga gagagccgtt atcgtctgtt tgtggatgta
      961 cagagtgata ttattgacac gcccgggcga cggatggtga tccccctggc cagtgcacgt
     1021 ctgctgtcag ataaagtctc ccgtgaactt tacccggtgg tgcatatcgg ggatgaaagc
     1081 tggcgcatga tgaccaccga tatggccagt gtgcctgtct ccgttatcgg ggaagaagtg
     1141 gctgatctca gccaccgcga aaatgacatc aaaaacgcca ttaacctgat gttctgggga
     1201 atataattaa gccagccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     1261 ctgctcccgg catccgctta cagacaagct gtgacccact caagtgggtc tcagttttag
     1321 agctagaaat agcaagttaa aataaggcta gtccgttatc aacttgaaaa agtggcaccg
     1381 agtcggtgct ttttttacgg tcatgtaaaa aaaaaccccg cccctgacag ggcggggttt
     1441 tttttcagtc aagcctctct tgacatcttt ccccgagata aatcggtaag ttgctgatta
     1501 aaaaactcca acatttctat cggcgccatg ccgatagcat cgctctccag gctgcgcgcg
     1561 tactgcgcca gcgccaggca tctctgcgcc agcatgtcgc agcccaggct gtagtcgctg
     1621 ctgacccggc tgctgaccgg ctcgggcacg ggcggtccag catggcggcc aggcaggccg
     1681 gacggcatgt tcatcagggc cgccgacagg gggcgcaaca ggcctgtcat gatgtcgatg
     1741 gacgcgttca tcaggcgcga gcggcgcagg ctgcccagcg gctgctgcgc aaaatggtgc
     1801 gccatcatca ggaaggtcag ttcatagcag ccctgataca gctgcatcag cgcgcgcgcc
     1861 ttctgatcgg tcaccacgct gcagtccgcg cgcgcttcca gcacgggatt cttcagcgcc
     1921 ggcagcgccg gttcgaacgg cttgtcgcag gccgaaaacc tgccggcgat ggcgcgcagc
     1981 cgctggaagt gcgaggaggc gaaatgcggc gaatcgacgg ccacgccttc gccctgttcc
     2041 gtgacgaaat cgatggcgaa caacgcgctg tcgcggtcgg atacttccag ctggtagccg
     2101 ggataggcgc ggttggtcag ttctttcagg aacaggtggt gctcgccgcc gcgcttgccg
     2161 ccgccgcttt cgaacaggcc gggcagctcg gccagggcct ggcggatcgc gatatagaag
     2221 gtggcgatgg atttgcccgg cgtgggaatg tagtcgggcc attcaaaacg cacgaagcgg
     2281 gccagcacgt gttcggaaaa tggttcgaac gcaaattccg tgtccaggcc gaaacgctgg
     2341 cgcgcctgct ggcccagcag cggggtaccg ctgtaaaacg gttcgccaag cgccatcaat
     2401 acattgttca ccaataagta gtgaatcatt tcttcatggg cgatttccag cagcgcgccg
     2461 cgcgtgccgc tgttgcggcg ccgatcctcg gcgccacacg ctaactccag ttccgcggga
     2521 agccagcgac cagactgcac caacgcggca ccttgcgcat aattcgggat gctatacgca
     2581 gcgtacagat actgcagcat caggcttaat tccaggtcaa tcgccttctt cagctcatct
     2641 atcagttcag ctttgccgcc aatcgcatga ggcgccgctg gttccggagc cgccgctttt
     2701 gcggccgcgg cggcttctac ttgggtcaga tatttcagaa acaagcggga tttaggcagc
     2761 gacagttcgc gagtgctggg catgtagtag cttttgtccc gattttgcgg gtcacacatc
     2821 tgccacatca ggcgtgaata cgtttcgcat ttgcactgat cagccaggct gaacacctta
     2881 tcggacataa aggggtaaac caattcgtag taactcatca catggcggta caagaaggca
     2941 tagtccactt gttctgctgg aacgtcgtct aagtcccagt catccggcag aacccgcacg
     3001 ccaagatact gtttcgcttt gccagttcca aggacaatgc gcgttgctcc cggtttacga
     3061 cctgtgaggg tcagcgtggt atagccgtgg cctaacggac tcggttgctg ctcgacggct
     3121 aagaggacgc catcttccgc ttcggccagt gacggaggtg cagccagttc accacggaaa
     3181 cgcgattgaa ccgtgatggt ctgcggaaac tgctcgtgtt tgttgcgatt cggtgcctcc
     3241 aggtacagct ggttagaatc gctttggagg acccaatcgg cttcatccca ctgtgcctgg
     3301 gctgaaccga gcgaaagaga acctggtgca gctcccgcat gctgtaatgg aacatcaaac
     3361 acgccatgat gacgccaata gtcgcgatac aactgttccg gaatacgggc gagtacggtc
     3421 cctgccccat cgtgcagcag cagatcgccc aatgcttgtt taccacccag cgcatgtgtc
     3481 ggatgctgct cggatacagc cccggcatca cgggtagtaa acgggattgc ggtaggcatg
     3541 ttgaaactca cgcgatccgc atgcactttc accagcaccg gccccaggct gccctggcgc
     3601 ggctgcagca ggcggccggc cggataggtg gccagctcgc cgcgccgcca caggccgatg
     3661 ctgccggcca ggtcgtagaa cacgggcgaa tcgggttttt gcggcgtcga catattgaac
     3721 aggcagtatt gcaccgtcag gcccagcacc tcgtcgtcgg ccagggcttg ctgcaaggca
     3781 tgcatactgg ccggcagcgg caggtccgga ttgaacagga aatgcgggtc ctgcttggcc
     3841 acggaaaact ggaacagcct ggagcggccg aattcctcgt ccaggaaatg cccgctgcgt
     3901 tccgtgatgt ggccgctgcc gagccagcgt accgagtgcg cctgcgcgat gtcggccgtg
     3961 aacagcgtgg gcgtattggg cgtggccagc ttgtcgctca aggtgaactg gcccgcgtag
     4021 atcagcgtgg tgtcgggctg cgccgggttg ttgtcgatcc accttgcgcg gttgaacgtc
     4081 gtgcgcaggt actcgttgta gtggccccac aggcccagct tggcgcccac cagcgcgtcc
     4141 tgggtatcga cctcgccatc acgcaactgg acgcccgtga tccgcgcgtt ttcccacgag
     4201 aaatggttgt tcccgcaaaa attatagcct gtcgcctggc tgaagatgcc gtccggatcg
     4261 ggcttgccct gtgcgttgaa gcggggggcg agctgtttca ggtgcgcatg gaattcggct
     4321 ggcggccggc tcaggtcgac agcctcgccc gccatcgata ccgcattcgt ggcgatatcg
     4381 atgttgccgt gcgtattgcg attccccgtc ggcacattgg cgcgggcaaa accccgaaaa
     4441 tgcaggcggg ggaagtcaag taggctcatt acgactctcc tctgtaaaag cgtccttgtc
     4501 ttccgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     4561 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     4621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     4681 ttcaataata ttgaaaaagg aagagtatgg ctaaaatgag aatatcaccg gaattgaaaa
     4741 aactgatcga aaaataccgc tgcgtaaaag atacggaagg aatgtctcct gctaaggtat
     4801 ataagctggt gggagaaaat gaaaacctat atttaaaaat gacggacagc cggtataaag
     4861 ggaccaccta tgatgtggaa cgggaaaagg acatgatgct atggctggaa ggaaagctgc
     4921 ctgttccaaa ggtcctgcac tttgaacggc atgatggctg gagcaatctg ctaatgagtg
     4981 aggccgatgg cgtcctttgc tcggaagagt atgaagatga acaaagccct gaaaagatta
     5041 tcgagctgta tgcggagtgc atcaggctct ttcactccat cgacatatcg gattgtccct
     5101 atacgaatag cttagacagc cgcttagccg aattggatta cttactgaat aacgatctgg
     5161 ccgatgtgga ttgcgaaaac tgggaagagg acactccatt taaagatccg cgcgagctgt
     5221 atgatttttt aaagacggaa aagcccgaag aggaacttgt cttttcccac ggcgacctgg
     5281 gagacagcaa catctttgtg aaagatggca aagtaagtgg ctttattgat cttgggagaa
     5341 gcggcagggc ggacaagtgg tatgacattg ccttctgcgt ccggtcgctc agggaggata
     5401 tcggggaaga acagtatgtc gagctatttt ttgacttact ggggatcaag cctgattggg
     5461 agaaaataaa atattatatt ttactggatg aattgtttta gctgtcagac caagtttact
     5521 ccggggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc
     5581 ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat
     5641 aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc
     5701 cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct
     5761 cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg
     5821 aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc
     5881 cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga
     5941 ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa
     6001 gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta
     6061 gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc
     6121 agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg
     6181 acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga
     6241 tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg
     6301 agtaaacttg gtctggcaga agaccatc
//
