LOCUS       sequence-140521-        2111 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Backbone vector with internal BsaI removed by site-directed 
            mutagenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2111)
  AUTHORS   Moore SJ, Lai HE, Kelwick RJ, Chee SM, Bell DJ, Polizzi KM, Freemont
            PS
  TITLE     EcoFlex: A Multifunctional MoClo Kit for E. coli Synthetic Biology.
  JOURNAL   ACS Synth Biol. 2016 May 2.
  PUBMED    27096716
REFERENCE   2  (bases 1 to 2111)
  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: "ACS Synth 
            Biol. 2016 May 2."
COMMENT     Sequence Label: pTU1
FEATURES             Location/Qualifiers
     source          1..2111
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(65..82)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(236..824)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(316..335)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(995..1786,1787..1855))
                     /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     1618..1637
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1856..1960)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2028..2046
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2050..2093)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(2096..2111,1..4)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
ORIGIN
        1 ctagaacgtc tcaatctcta tagagaccca cggtctcatg ttttagagag acgctgcagg
       61 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
      121 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
      181 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
      241 acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
      301 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
      361 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
      421 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
      481 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
      541 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
      601 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      661 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      721 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      781 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      841 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      901 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      961 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     1021 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     1081 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagagc
     1141 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     1201 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     1261 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     1321 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     1381 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     1441 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     1501 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1561 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1621 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1681 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1741 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1801 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1861 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1921 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1981 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     2041 cgaggcagaa tttcagataa aaaaaatcct tagctttcgc taaggatgat ttctggaatt
     2101 cgcggccgct t
//
