LOCUS       sequence-363161-        3251 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For scarless integration into the chromosome, HAIB plasmid can be 
            usedin place of the HA, INS and HB modules..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3251)
  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 3251)
  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_HAIB
FEATURES             Location/Qualifiers
     source          1..3251
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(466..1054)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(546..565)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    1430..1451
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1466..1471,1472..1489,1490..1496)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1504..1520
                     /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     1509..1531
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1528..1544
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1528..1544
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1540..1863
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCTLSTICS
                     DAA"
     misc_feature    1557..1613
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1614..1631)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1614..1630)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1623..1645)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(1839..1858)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     CDS             2205..2510
                     /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     2389..2408
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     primer_bind     complement(2672..2690)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2758..2862
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2863..3251,1..406)
                     /codon_start=1
                     /gene="aphA-3"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /translation="MAKMRISPELKKLIEKYRCVKDTEGMSPAKVYKLVGENENLYLKM
                     TDSRYKGTTYDVEREKDMMLWLEGKLPVPKVLHFERHDGWSNLLMSEADGVLCSEEYED
                     EQSPEKIIELYAECIRLFHSIDISDCPYTNSLDSRLAELDYLLNNDLADVDCENWEEDT
                     PFKDPRELYDFLKTEKPEEELVFSHGDLGDSNIFVKDGKVSGFIDLGRSGRADKWYDIA
                     FCVRSLREDIGEEQYVELFFDLLGIKPDWEKIKYYILLDELF"
ORIGIN
        1 tccctatacg aatagcttag acagccgctt agccgaattg gattacttac tgaataacga
       61 tctggccgat gtggattgcg aaaactggga agaggacact ccatttaaag atccgcgcga
      121 gctgtatgat tttttaaaga cggaaaagcc cgaagaggaa cttgtctttt cccacggcga
      181 cctgggagac agcaacatct ttgtgaaaga tggcaaagta agtggcttta ttgatcttgg
      241 gagaagcggc agggcggaca agtggtatga cattgccttc tgcgtccggt cgctcaggga
      301 ggatatcggg gaagaacagt atgtcgagct attttttgac ttactgggga tcaagcctga
      361 ttgggagaaa ataaaatatt atattttact ggatgaattg ttttagctgt cagaccaagt
      421 ttactccggg gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
      481 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
      541 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
      601 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
      661 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
      721 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
      781 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
      841 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
      901 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
      961 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     1021 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     1081 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     1141 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     1201 atatgagtaa acttggtctg gcgaaaaaac cccgccgaag cggggttttt tgcggtctga
     1261 ctgcagaaga cagccgtcga gtacattata ccgtggcaca tttatctgtc caactcctat
     1321 atcggcataa accaaggtcc aaacggtgac ggccgttgag acccagtggt atgggcagct
     1381 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt
     1441 agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg
     1501 gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccaagc
     1561 ttgcatgcct gcaggtcgac tctagaggat ccccgggtac cgagctcgaa ttcactggcc
     1621 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     1681 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     1741 caacagttgc gcagcctgaa tggcgaatgg cgcctgatgc ggtattttct ccttacgcat
     1801 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     1861 tagtcgcgtg gatccggctt actaaaagcc agataacagt atgcgtattt gcgcgctgat
     1921 ttttgcggta taagaatata tactgatatg tatacccgaa gtatgtcaaa aagaggtatg
     1981 ctatgaagca gcgtattaca gtgacagttg acagcgacag ctatcagttg ctcaaggcat
     2041 atatgatgtc aatatctccg gtctggtaag cacaaccatg cagaatgaag cccgtcgtct
     2101 gcgtgccgaa cgctggaaag cggaaaatca ggaagggatg gctgaggtcg cccggtttat
     2161 tgaaatgaac ggctcttttg ctgacgagaa caggggctgg tgaaatgcag tttaaggttt
     2221 acacctataa aagagagagc cgttatcgtc tgtttgtgga tgtacagagt gatattattg
     2281 acacgcccgg gcgacggatg gtgatccccc tggccagtgc acgtctgctg tcagataaag
     2341 tctcccgtga actttacccg gtggtgcata tcggggatga aagctggcgc atgatgacca
     2401 ccgatatggc cagtgtgcct gtctccgtta tcggggaaga agtggctgat ctcagccacc
     2461 gcgaaaatga catcaaaaac gccattaacc tgatgttctg gggaatataa ttaagccagc
     2521 cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg
     2581 cttacagaca agctgtgacc cactcaagtg ggtctcacgc tccgtcaccg tttggacctt
     2641 ggttcagtgg tctgtcttcc gagtcgaaag ggcctcgtga tacgcctatt tttataggtt
     2701 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     2761 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     2821 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatggctaa aatgagaata
     2881 tcaccggaat tgaaaaaact gatcgaaaaa taccgctgcg taaaagatac ggaaggaatg
     2941 tctcctgcta aggtatataa gctggtggga gaaaatgaaa acctatattt aaaaatgacg
     3001 gacagccggt ataaagggac cacctatgat gtggaacggg aaaaggacat gatgctatgg
     3061 ctggaaggaa agctgcctgt tccaaaggtc ctgcactttg aacggcatga tggctggagc
     3121 aatctgctaa tgagtgaggc cgatggcgtc ctttgctcgg aagagtatga agatgaacaa
     3181 agccctgaaa agattatcga gctgtatgcg gagtgcatca ggctctttca ctccatcgac
     3241 atatcggatt g
//
