LOCUS       sequence-363178-        5414 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pSwap plasmid part containing the T2 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 5414)
  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 5414)
  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_T2
FEATURES             Location/Qualifiers
     source          1..5414
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(523..1111)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(603..622)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1428..1446
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1453..1471
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1555..1576
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1591..1596,1597..1614,1615..1621)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1629..1645
                     /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     1634..1656
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1653..1669
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1653..1669
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     misc_feature    1682..1738
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1739..1756)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1739..1755)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1748..1770)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1964..1983)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             2330..2635
                     /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="MQFKVYTYKRESR*RLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTGMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     misc_RNA        2743..2818
                     /label=gRNA scaffold
                     /note="guide RNA scaffold for the Streptococcus pyogenes 
                     CRISPR/Cas9 system"
     regulatory      4168..4177
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(4892..4910)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4978..5082
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5083..5414,1..463)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tatcgagctg tatgcggagt gcatcaggct ctttcactcc atcgacatat cggattgtcc
       61 ctatacgaat agcttagaca gccgcttagc cgaattggat tacttactga ataacgatct
      121 ggccgatgtg gattgcgaaa actgggaaga ggacactcca tttaaagatc cgcgcgagct
      181 gtatgatttt ttaaagacgg aaaagcccga agaggaactt gtcttttccc acggcgacct
      241 gggagacagc aacatctttg tgaaagatgg caaagtaagt ggctttattg atcttgggag
      301 aagcggcagg gcggacaagt ggtatgacat tgccttctgc gtccggtcgc tcagggagga
      361 tatcggggaa gaacagtatg tcgagctatt ttttgactta ctggggatca agcctgattg
      421 ggagaaaata aaatattata ttttactgga tgaattgttt tagctgtcag accaagttta
      481 ctccggggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
      541 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
      601 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
      661 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
      721 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
      781 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
      841 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
      901 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
      961 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     1021 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     1081 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     1141 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     1201 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     1261 tgagtaaact tggtctggcg aaaaaacccc gccgaagcgg ggttttttgc ggtctgactg
     1321 cagaagacca gtcctgagtt agctagatat cgatacatta taacagttcc ttctgcaaat
     1381 gtcaactcct atggtgctga tcggctttac gtgcgctact agtatgctcc ctatcagtga
     1441 tagagattga catccctatc agtgatagag atactgagca ctgagaccca gtggtatggg
     1501 cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt
     1561 gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt
     1621 gtgtggaatt gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc
     1681 caagcttgca tgcctgcagg tcgactctag aggatccccg ggtaccgagc tcgaattcac
     1741 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     1801 ttgcaggaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     1861 cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat tttctcctta
     1921 cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc tgctctgatg
     1981 ccgcatagtc gcgtggatcc ggcttactaa aagccagata acagtatgcg tatttgcgcg
     2041 ctgatttttg cggtataaga atatatactg atatgtatac ccgaagtatg tcaaaaagag
     2101 gtatgctatg aagcagcgta ttacagtgac agttgacagc gacagctatc agttgctcaa
     2161 ggcatatatg atgtcaatat ctccggtctg gtaagcacaa ccatgcagaa tgaagcccgt
     2221 cgtctgcgtg ccgaacgctg gaaagcggaa aatcaggaag ggatggctga ggtcgcccgg
     2281 tttattgaaa tgaacggctc ttttgctgac gagaacaggg gctggtgaaa tgcagtttaa
     2341 ggtttacacc tataaaagag agagccgtta gcgtctgttt gtggatgtac agagtgatat
     2401 tattgacacg cccgggcgac ggatggtgat ccccctggcc agtgcacgtc tgctgtcaga
     2461 taaagtctcc cgtgaacttt acccggtggt gcatatcggg gatgaaagct ggcgcatgat
     2521 gaccaccggt atggccagtg tgcctgtctc cgttatcggg gaagaagtgg ctgatctcag
     2581 ccaccgcgaa aatgacatca aaaacgccat taacctgatg ttctggggaa tataattaag
     2641 ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     2701 atccgcttac agacaagctg tgacccactc aagtgggtct cagttttaga gctagaaata
     2761 gcaagttaaa ataaggctag tccgttatca acttgaaaaa gtggcaccga gtcggtgctt
     2821 tttttacggt catgtaaaaa aaaaccccgc ccctgacagg gcggggtttt ttttcagtct
     2881 gactgtatct ctttgggcga aaaaaccccg ccgaagcggg gttttttgcg ttaggtgttg
     2941 caagacgtaa agacgctgtc cggaatgtcg ctgacaaaca ggttgggaaa gtcgcgtacc
     3001 gacaggtgct gcgggtcatt gccggtgacc atgcgcagca ggcggttgcc accggcctcc
     3061 aggtaaaaga ccgatggcgg cttgcccgcc cgctcgacta cccaggcgtc cgcctcccgg
     3121 cccagcacgg tatggcggcc gtcgtgacgg gcaccgccgt ccagcagcac cgcctgcggc
     3181 aggaacagct catggaacgg ggtcacctca tcggccagcg ccgtgccctg cagctgctcg
     3241 ccagccgcct ccgcttgcct tgcgtaggcc actttctgct tgcggctttg ttcgcgcctg
     3301 gcgtcgccga tttccgacat ccacagcagg tgtccatgtt ccttttccga ccaggggttg
     3361 aacaggccgt cgatgcggca gatattgcgc gcatagtcga accagcaata gccggaggtg
     3421 acctggtcat ccgcctgcat cggcgtccag taggaaacat aggcgctgct ccattgcatc
     3481 ggcagcagcg gcgggctgac gtgtgtcggc attttaattt tcctccgtaa aagcgctcat
     3541 gcgcgctcgg tcgaggaagt gatgaaggta tcgttggcag cttcttcttc cgtccgttga
     3601 tcgaggcgct gcagcaacag gccgctggcg tgctgggcgc tgatcaggct gccttccatc
     3661 cagccgcaat gcgcggtata ggcatccgag caggagatga tgccgtcccg gtgcagcagg
     3721 atggccgggt ggtcggcttc cggctccacg caaaactcga cgccatgcgg ccagaacttg
     3781 tggaagtgcg ccttgatctg cggcagaggc tggccatcga gcggcaggac ttcttccagg
     3841 tggctgcgga cacgctccag gtatacgtcc tcaccaagct ccaggctgtc ccgccagtag
     3901 gtggcgcttt tgctgtcggt gtagaacagc acgtatttgt cgctcttgaa gtagatcttg
     3961 cgcaggggat ttgccgccat cagcaccttg tcggtcagcc ccagcgcatc ccaccaggct
     4021 gtgtcgaacg tgaagaatcc cttgaacagg ggcagcgagt cgtattggaa cggactccag
     4081 gcgttcggga aatccaggtt caggcgcggc atggcggacg gcgggatggc catcaccaga
     4141 tggcgcgtgc ggtgcatctg cgtgtcgcca tggtggctga aggcgagcac atggtcggcg
     4201 cccgattttt caacggacag caagcggtgt tcgagcctga attccacccc ggcgacctgg
     4261 gcctggtgct gcaactggca cagcaattcg tggtagccgt cggtggcgta gtgccactgg
     4321 ttggcggcgt tttccgtaaa gtgctgcgtt tccgggtgct tcttgatgat gtcgtaggcc
     4381 atggccgccg acaccatcgg cagcagcagg gcgtcatagc cggtcgcctt gatgatgtgg
     4441 ctggccttgg cggcgcccag gtaatggctg acgaactcga ggaaggagtc gttcggatgc
     4501 attttcagca tcgggctcag ctcatccagc gttgccttca gctcctccag cacgctatcg
     4561 tgggagatga cctcggtgaa cgggtagacc gcatgcggca agccgctgcc ctgcatgagt
     4621 tgctggaaat gcggatgcag ctgcggcgag tagcgggcgg cgccgagttc ggcgatttcc
     4681 tcgccatcta ttttccgcga ctggatgcgg ccgcctacgg tggcattcag gtcgaatacg
     4741 cggatgcgca ggttcctgcc ggcggcggcg tcgatcaggt tgttggcgca agtcaagcct
     4801 cctatgccgg cgccaacgat gcaaatgtca gaatacgtgc tcattgtgcg gctcctccgt
     4861 aaaagcgtct tggtcttcgc gagacgaaag ggcctcgtga tacgcctatt tttataggtt
     4921 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     4981 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5041 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata
     5101 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
     5161 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
     5221 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
     5281 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
     5341 aatctgctaa tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
     5401 agccctgaaa agat
//
