LOCUS       sequence-447511-        2516 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Level 1 Destination vector - Position D.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2516)
  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 2516)
  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-D-lacZ
FEATURES             Location/Qualifiers
     source          1..2516
                     /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)"
     primer_bind     complement(464..481)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(635..1223)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(715..734)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1394..2185,2186..2254))
                     /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     2017..2036
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2255..2359)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2427..2445
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2449..2492)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2495..2514
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
ORIGIN
        1 cgtctcagaa gctatagaga cctctagaca 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 ggtctcatgt tttagagaga cgctgcaggc ttcctcgctc actgactcgc
      481 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
      541 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
      601 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
      661 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
      721 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
      781 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
      841 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
      901 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
      961 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     1021 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     1081 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     1141 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     1201 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     1261 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     1321 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     1381 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     1441 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     1501 taccatctgg ccccagtgct gcaatgatac cgcgagagcc acgctcaccg gctccagatt
     1561 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     1621 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     1681 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     1741 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     1801 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     1861 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     1921 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     1981 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     2041 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     2101 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     2161 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     2221 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     2281 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     2341 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     2401 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa
     2461 aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt ctagaa
//
