LOCUS       sequence-218371-        2599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Upstream homology arm, amyE encodes _-amylase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2599)
  AUTHORS   Wicke N, Radford D, Verrone V, Wipat A, French CE
  TITLE     BacilloFlex: A modular DNA assembly toolkit for Bacillus subtilis 
            synthetic biology
  JOURNAL   bioRxiv 185108
REFERENCE   2  (bases 1 to 2599)
  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 
            185108"
COMMENT     Sequence Label: pTU1-A-amyEU
FEATURES             Location/Qualifiers
     source          1..2599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(547..564)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(718..1306)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(798..817)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1477..2268,2269..2337))
                     /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     2100..2119
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2338..2442)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2510..2528
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2532..2575)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2578..2597
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
ORIGIN
        1 cgtctcaatc ttttgcaaaa cgattcaaaa cctctttact gccgttattc gctggatttt
       61 tattgctgtt tcatttggtt ctggcaggac cggcggctgc gagtgctgaa acggcgaaca
      121 aatcgaatga gcttacagca ccgtcgatca aaagcggaac cattcttcat gcatggaatt
      181 ggtcgttcaa tacgttaaaa cacaatatga aggatattca tgatgcagga tatacagcca
      241 ttcagacatc tccgattaac caagtaaagg aagggaatca aggagataaa agcatgtcga
      301 actggtactg gctgtatcag ccgacatcgt atcaaattgg caaccgttac ttaggtactg
      361 aacaagaatt taaagaaatg tgtgcagccg ctgaagaata tggcataaag gtcattgttg
      421 acgcggtcat caatcatacc accagtgatt atgccgcgat ttccaatgag gttaagagta
      481 ttccaaactg gacacatgga aacacacaaa ttaaaaactg gtcttgccag agacgctgca
      541 ggcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
      601 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
      661 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
      721 ccacaggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
      781 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
      841 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
      901 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
      961 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     1021 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     1081 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     1141 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     1201 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     1261 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     1321 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     1381 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     1441 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     1501 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     1561 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     1621 gccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     1681 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     1741 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     1801 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     1861 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     1921 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     1981 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     2041 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     2101 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     2161 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     2221 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     2281 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     2341 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     2401 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     2461 gccacctgac gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat
     2521 cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg atttctggaa
     2581 ttcgcggccg cttctagaa
//
