LOCUS       sequence-133795-        2506 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 2 Destination vector - Position A - 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-A-lacZ
FEATURES             Location/Qualifiers
     source          1..2506
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    36..52
                     /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     41..63
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     60..76
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     60..76
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             85..96
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     terminator      460..517
                     /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(541..558)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(712..1300)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(792..811)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1483..1577)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1598..2257)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2172..2191
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2419..2437
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2441..2484)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    join(2487..2506,1..2)
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
ORIGIN
        1 aggtctcact atatctagag acgtctagac atatgttgtg agcggataac aatttcacac
       61 aggaaacagc tatgaccatg gtacaaagag gagaaaggac atgggtgacc cgaaactgct
      121 gctggaagtt ccgcacgcta tctcttcttc tccgggtaac tctctggctg ttgttctgca
      181 acgtcgtgac tgggaaaacc cgggtgttac ccagctgaac cgtctggctg ctcacccgcc
      241 gttcgcttct tggcgtaact ctgaagaagc tcgtaccgac cgtccgtccc agcagctgcg
      301 ttctctgatc cgtctgctga ccaaaccgga acgtaaactg tcttggctgc tgccgccgct
      361 gtctaacaac taacgcaaaa aaccccgccc ctgacagggc ggggtttttt cgcactagtg
      421 gatccgcatg ccgtctcatt aggtacagag accctgcagt ccggcaaaaa agggcaaggt
      481 gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta tgcaggcttc
      541 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
      601 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
      661 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccacag
      721 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
      781 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
      841 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
      901 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
      961 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     1021 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     1081 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     1141 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     1201 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     1261 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     1321 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     1381 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1441 aagtatatat gagtaaactt ggtctgacag ctcgaggctt ggattctcac caataaaaaa
     1501 cgcccggcgg caaccgagcg ttctgaacaa atccagatgg agttctgagg tcattactgg
     1561 atctatcaac aggagtccaa gcgagctcga tatcaaatta cgccccgccc tgccactcat
     1621 cgcagtactg ttgtaattca ttaagcattc tgccgacatg gaagccatca caaacggcat
     1681 gatgaacctg aatcgccagc ggcatcagca ccttgtcgcc ttgcgtataa tatttgccca
     1741 tggtgaaaac gggggcgaag aagttgtcca tattggccac gtttaaatca aaactggtga
     1801 aactcaccca gggattggct gacacgaaaa acatattctc aataaaccct ttagggaaat
     1861 aggccaggtt ttcaccgtaa cacgccacat cttgcgaata tatgtgtaga aactgccgga
     1921 aatcgtcgtg gtattcactc cagagcgatg aaaacgtttc agtttgctca tggaaaacgg
     1981 tgtaacaagg gtgaacacta tcccatatca ccagctcacc gtctttcatt gccatacgaa
     2041 attccggatg agcattcatc aggcgggcaa gaatgtgaat aaaggccgga taaaacttgt
     2101 gcttattttt ctttacggtc tttaaaaagg ccgtaatatc cagctgaacg gtctggttat
     2161 aggtacattg agcaactgac tgaaatgcct caaaatgttc tttacgatgc cattgggata
     2221 tatcaacggt ggtatatcca gtgatttttt tctccatttt agcttcctta gctcctgaaa
     2281 atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg tgaaagttgg
     2341 aacctcttac gtgcccgatc aactcgagtg ccacctgacg tctaagaaac cattattatc
     2401 atgacattaa cctataaaaa taggcgtatc acgaggcaga atttcagata aaaaaaatcc
     2461 ttagctttcg ctaaggatga tttctggaat tcgcggccgc ttctag
//
