LOCUS       sequence-406998-        2116 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid for ORBIT in E. coli. Orthogonal attP site 
            (#5)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2116)
  AUTHORS   Saunders SH, Ahmed AM
  TITLE     ORBIT for E. coli: Kilobase-scale oligonucleotide recombineering at 
            high throughput and high efficiency
  JOURNAL   bioRxiv 2023
REFERENCE   2  (bases 1 to 2116)
  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: "bioRxiv 
            2023"
COMMENT     Sequence Label: pInt_attP5_ampR
FEATURES             Location/Qualifiers
     source          1..2116
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..48
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     protein_bind    complement(69..116)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     primer_bind     complement(286..304)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        372..476
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(477..545,546..1337)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(695..714)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1619..2007
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(2051..2084)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
ORIGIN
        1 ggtttgtctg gtcaaccacc gcgctctcag tggtgtacgg tacaaaccgt gtaggctgga
       61 gctgcttcga agttcctata ctttctagag aataggaact tcggaatagg aacttcaaga
      121 tcccccacgc tgccgcaagc actcagggcg caagggctgc taaaggaagc ggaacacgta
      181 gaaagccagt ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg
      241 gacaagggaa aacgcaagcg caaagagaaa gcaggtaacg aaagggcctc gtgatacgcc
      301 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
      361 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
      421 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
      481 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
      541 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
      601 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
      661 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg
      721 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
      781 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
      841 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
      901 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
      961 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     1021 cagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     1081 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     1141 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     1201 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     1261 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     1321 tgattaagca ttggtaactg tcagaccaag tttactcatc caattattga acacccttcg
     1381 gggtgttttt ttgtttctgg tctcccttcg aaatgaccga ccaagcgacg cccaacctgc
     1441 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     1501 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     1561 accccagctt caaaagcgct ctaaggagga tattcatatg gaccatggct aattcccatg
     1621 tcagccgtta agtgttcctg tgtcactcaa aattgctttg agaggctcta agggcttctc
     1681 agtgcgttac atccctggct tgttgtccac aaccgttaaa ccttaaaagc tttaaaagcc
     1741 ttatatattc ttttttttct tataaaactt aaaaccttag aggctattta agttgctgat
     1801 ttatattaat tttattgttc aaacatgaga gcttagtacg tgaaacatga gagcttagta
     1861 cgttagccat gagagcttag tacgttagcc atgagggttt agttcgttaa acatgagagc
     1921 ttagtacgtt aaacatgaga gcttagtacg tgaaacatga gagcttagta cgtactatca
     1981 acaggttgaa ctgctgatct tcagatcctc tacgccggac gcatcgtggc cggatcttgc
     2041 ggccgcaaaa gaagttccta tactttctag agaataggaa cttcggaata ggaactatta
     2101 cacgtcttga gcgatt
//
