LOCUS       sequence-140543-         778 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Level 2 Destination vector - Position B - Assembly of 4-5 TU's from 
            Level 1-ABCD or ABCDE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 778)
  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 778)
  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: pTU2-B-lacZ
FEATURES             Location/Qualifiers
     source          1..778
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     terminator      complement(22..65)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    68..89
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    123..139
                     /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)."
     primer_bind     128..150
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     147..163
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     147..163
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             172..183
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      547..604
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(628..645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ataggcgtat cacgaggcag aatttcagat aaaaaaaatc cttagctttc gctaaggatg
       61 atttctggaa ttcgcggccg cttctagagg tctcagtaca tctagagacg tctagacata
      121 tgttgtgagc ggataacaat ttcacacagg aaacagctat gaccatggta caaagaggag
      181 aaaggacatg ggtgacccga aactgctgct ggaagttccg cacgctatct cttcttctcc
      241 gggtaactct ctggctgttg ttctgcaacg tcgtgactgg gaaaacccgg gtgttaccca
      301 gctgaaccgt ctggctgctc acccgccgtt cgcttcttgg cgtaactctg aagaagctcg
      361 taccgaccgt ccgtcccagc agctgcgttc tctgatccgt ctgctgacca aaccggaacg
      421 taaactgtct tggctgctgc cgccgctgtc taacaactaa cgcaaaaaac cccgcccctg
      481 acagggcggg gttttttcgc actagtggat ccgcatgccg tctcattagg gacagagacc
      541 ctgcagtccg gcaaaaaagg gcaaggtgtc accaccctgc cctttttctt taaaaccgaa
      601 aagattactt cgcgttatgc aggcttcctc gctcactgac tcgctgcgct cggtcgttcg
      661 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      721 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaa
//
