LOCUS       sequence-121749-        3569 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  source of TpR FRT cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3569)
  AUTHORS   Choi KH, Gaynor JB, White KG, Lopez C, Bosio CM, Karkhoff-Schweizer 
            RR, Schweizer HP
  TITLE     A Tn7-based broad-range bacterial cloning and expression system.
  JOURNAL   Nat Methods. 2005 Jun;2(6):443-8.
  PUBMED    15908923
REFERENCE   2  (bases 1 to 3569)
  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 
            Methods."; date: "2005-06"; volume: "2(6)"; pages: "443-8"
COMMENT     Sequence Label: pFTP1
FEATURES             Location/Qualifiers
     source          1..3569
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(29..51)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     151..170
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     364..386
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     378..395
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     379..395
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    464..511
                     /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)."
     CDS             complement(573..809)
                     /codon_start=1
                     /gene="dhfrIIb from plasmid R388"
                     /product="E. coli plasmid-associated dihydrofolate 
                     reductase"
                     /label=TpR
                     /note="confers resistance to trimethoprim"
                     /translation="MGQSSDEANAPVAGQFALPLSATFGLGDRVRKKSGAAWQGQVVGW
                     YCTKLTPEGYAVESESHPGSVQIYPVAALERVA"
     promoter        complement(join(1135..1140,1141..1157,1158..1163))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    1215..1262
                     /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(1348..1364)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1348..1364)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1361..1383)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1372..1388
                     /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)."
     promoter        complement(join(1396..1402,1403..1420,1421..1426))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1441..1462
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1579..1596)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1750..2338)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1830..1849)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2509..3300,3301..3369))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3132..3151
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3370..3474)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3542..3560
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa gcttgcatgc ctgcaggtcg
      421 actctagagg atccccgggt accgagctcg aattggggat cttgaagtac ctattccgaa
      481 gttcctattc tctagaaagt ataggaactt cagagcgctt ttgaagctga tgtgcttaaa
      541 aacttactca atggaataat tctagataat tcttaggcca cacgttcaag tgcagccaca
      601 ggataaattt gcactgagcc tgggtgggat tcggactcga ccgcatagcc ttcaggagtg
      661 agttttgtgc aataccaacc gacgacttga ccctgccaag cggcaccaga tttcttgcgt
      721 acgcgatccc ctaagccaaa ggtggcactc aggggaagcg caaactgccc tgcaacggga
      781 gcgttggctt catcgctact ttgacccatg tcgaatcctt cttgtgaatc tattatggcg
      841 acaagcaaat cgagctctga ctgcctaccc cacaacaact atcagaaagc accagcacaa
      901 cggctgccta actttgtttt agggcgactg ccctgctgcg taacatcgtt gctgctccat
      961 aacatcaaac atcgacccac ggcgtaacgc gcttgctgct tggatgcccg aggcatagac
     1021 tgtacaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt accaccgctg
     1081 cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca ttacagttta
     1141 cgaaccgaac aggcttatgt caactgggtt cgtgaattat ccattgctgt tgacaaaggg
     1201 aatcagggga tcttgaagtt cctattccga agttcctatt ctctagaaag tataggaact
     1261 tcagagcgct tttgaagcta attcgagctc ggtacccggg gatcctctag agtcgacctg
     1321 caggcatgca agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc
     1381 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     1441 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     1501 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat
     1561 tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     1621 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     1681 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     1741 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     1801 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     1861 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     1921 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     1981 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     2041 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     2101 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     2161 gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa
     2221 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     2281 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     2341 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     2401 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     2461 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     2521 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     2581 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     2641 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     2701 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     2761 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     2821 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     2881 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     2941 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     3001 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     3061 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     3121 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     3181 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     3241 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     3301 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     3361 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     3421 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     3481 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     3541 aaataggcgt atcacgaggc cctttcgtc
//
