LOCUS       sequence-447522-        2506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 Destination vector - Position D - Assembly of 4-5 TU's from 
            Level 1-ABCD or ABCDE.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2506)
  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 2506)
  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-D-lacZ
FEATURES             Location/Qualifiers
     source          1..2506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    35..51
                     /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     40..62
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     59..75
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     59..75
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             84..95
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      459..516
                     /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(540..557)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(711..1299)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(791..810)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1482..1576)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1597..2256)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2171..2190
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2418..2436
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2440..2483)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(2486..2506,1)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 ggtctcatcg aatctagaga cgtctagaca tatgttgtga gcggataaca atttcacaca
       61 ggaaacagct atgaccatgg tacaaagagg agaaaggaca tgggtgaccc gaaactgctg
      121 ctggaagttc cgcacgctat ctcttcttct ccgggtaact ctctggctgt tgttctgcaa
      181 cgtcgtgact gggaaaaccc gggtgttacc cagctgaacc gtctggctgc tcacccgccg
      241 ttcgcttctt ggcgtaactc tgaagaagct cgtaccgacc gtccgtccca gcagctgcgt
      301 tctctgatcc gtctgctgac caaaccggaa cgtaaactgt cttggctgct gccgccgctg
      361 tctaacaact aacgcaaaaa accccgcccc tgacagggcg gggttttttc gcactagtgg
      421 atccgcatgc cgtctcatta gtgttagaga ccctgcagtc cggcaaaaaa gggcaaggtg
      481 tcaccaccct gccctttttc tttaaaaccg aaaagattac ttcgcgttat gcaggcttcc
      541 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
      601 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
      661 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccacagg
      721 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
      781 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
      841 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
      901 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
      961 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     1021 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     1081 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     1141 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     1201 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     1261 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     1321 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     1381 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     1441 agtatatatg agtaaacttg gtctgacagc tcgaggcttg gattctcacc aataaaaaac
     1501 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
     1561 tctatcaaca ggagtccaag cgagctcgat atcaaattac gccccgccct gccactcatc
     1621 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     1681 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     1741 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     1801 actcacccag ggattggctg acacgaaaaa catattctca ataaaccctt tagggaaata
     1861 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     1921 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     1981 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacgaaa
     2041 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     2101 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     2161 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     2221 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     2281 tctcgataac tcaaaaaata cgcccggtag tgatcttatt tcattatggt gaaagttgga
     2341 acctcttacg tgcccgatca actcgagtgc cacctgacgt ctaagaaacc attattatca
     2401 tgacattaac ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct
     2461 tagctttcgc taaggatgat ttctggaatt cgcggccgct tctaga
//
