LOCUS       sequence-407000-        1923 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integrating plasmid for ORBIT in E. coli. Orthogonal attP site 
            (#3)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1923)
  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 1923)
  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_attP3_chlorR
FEATURES             Location/Qualifiers
     source          1..1923
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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)."
     promoter        375..477
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             478..1137
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(544..563)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      1426..1814
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     protein_bind    complement(1858..1891)
                     /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 gcgcactcag 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 gcaggtatac ctgtgacgga agatcacttc
      301 gcagaataaa taaatcctgg tgtccctgtt gataccggga agccctgggc caacttttgg
      361 cgaaaatgag acgttgatcg gcacgtaaga ggttccaact ttcaccataa tgaaataaga
      421 tcactaccgg gcgtattttt tgagttatcg agattttcag gagctaagga agctaaaatg
      481 gagaaaaaaa tcactggata taccaccgtt gatatatccc aatggcatcg taaagaacat
      541 tttgaggcat ttcagtcagt tgctcaatgt acctataacc agaccgttca gctggatatt
      601 acggcctttt taaagaccgt aaagaaaaat aagcacaagt tttatccggc ctttattcac
      661 attcttgccc gcctgatgaa tgctcatccg gaattccgta tggcaatgaa agacggtgag
      721 ctggtgatat gggatagtgt tcacccttgt tacaccgttt tccatgagca aactgaaacg
      781 ttttcatcgc tctggagtga ataccacgac gatttccggc agtttctaca catatattcg
      841 caagatgtgg cgtgttacgg tgaaaacctg gcctatttcc ctaaagggtt tattgagaat
      901 atgtttttcg tctcagccaa tccctgggtg agtttcacca gttttgattt aaacgtggcc
      961 aatatggaca acttcttcgc ccccgttttc accatgggca aatattatac gcaaggcgac
     1021 aaggtgctga tgccgctggc gattcaggtt catcatgccg tttgtgatgg cttccatgtc
     1081 ggcagaatgc ttaatgaatt acaacagtac tgcgatgagt ggcagggcgg ggcgtaattt
     1141 ttttaaggca gttattggtg cccttaccaa ttattgaaca cccttcgggg tgtttttttg
     1201 tttctggtct cccttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     1261 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     1321 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccagcttcaa
     1381 aagcgctcta aggaggatat tcatatggac catggctaat tcccatgtca gccgttaagt
     1441 gttcctgtgt cactcaaaat tgctttgaga ggctctaagg gcttctcagt gcgttacatc
     1501 cctggcttgt tgtccacaac cgttaaacct taaaagcttt aaaagcctta tatattcttt
     1561 tttttcttat aaaacttaaa accttagagg ctatttaagt tgctgattta tattaatttt
     1621 attgttcaaa catgagagct tagtacgtga aacatgagag cttagtacgt tagccatgag
     1681 agcttagtac gttagccatg agggtttagt tcgttaaaca tgagagctta gtacgttaaa
     1741 catgagagct tagtacgtga aacatgagag cttagtacgt actatcaaca ggttgaactg
     1801 ctgatcttca gatcctctac gccggacgca tcgtggccgg atcttgcggc cgcaaaagaa
     1861 gttcctatac tttctagaga ataggaactt cggaatagga actattacac gtcttgagcg
     1921 att
//
