LOCUS       sequence-114495-        2731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Destination Vector: (based on pSB1K3)  [E:LacZa:F].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2731)
  AUTHORS   Iverson SV, Haddock TL, Beal J, Densmore DM
  TITLE     CIDAR MoClo: Improved MoClo Assembly Standard and New E. coli Part 
            Library Enable Rapid Combinatorial Design for Synthetic and 
            Traditional Biology.
  JOURNAL   ACS Synth Biol. 2016 Jan 15;5(1):99-103. doi: 
            10.1021/acssynbio.5b00124. Epub 2015 Nov 4.
  PUBMED    26479688
REFERENCE   2  (bases 1 to 2731)
  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"; doi: 
            "10.1021/acssynbio.5b00124"; journalName: "ACS Synth Biol"; date: 
            "2016-01-15- 15"; volume: "5"; issue: "1"; pages: "99-103"
COMMENT     Sequence Label: DVK_EF
FEATURES             Location/Qualifiers
     source          1..2731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     198..220
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     212..229
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     213..229
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    230..286
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(299..315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(299..315)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(312..334)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    323..339
                     /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)."
     promoter        complement(join(347..353,354..371,372..377))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    392..413
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    513..533
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      534..591
                     /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(615..632)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(786..1374)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(866..885)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1607..2422)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1607..2422)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2330..2349
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2414..2433
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2627..2645
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2649..2692)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2695..2716
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(2711..2731,1..303))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TSL"
ORIGIN
        1 gcttagagac ctgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
       61 ccgcatcagg cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
      121 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
      181 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct
      241 cggtacccgg ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt
      301 catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg
      361 gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt
      421 tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg
      481 gccaacgcgc gggggtctct cgctatgtct tctactagta gcggccgctg cagtccggca
      541 aaaaagggca aggtgtcacc accctgccct ttttctttaa aaccgaaaag attacttcgc
      601 gttatgcagg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
      661 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
      721 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
      781 gcgtttttcc acaggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
      841 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
      901 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
      961 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     1021 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     1081 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     1141 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     1201 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     1261 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     1321 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     1381 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     1441 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     1501 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagctcgag tcccgtcaag
     1561 tcagcgtaat gctctgccag tgttacaacc aattaaccaa ttctgattag aaaaactcat
     1621 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     1681 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     1741 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     1801 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     1861 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     1921 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgagac
     1981 gaaatacgcg atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca
     2041 ggaacactgc cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct
     2101 ggaatgctgt tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga
     2161 taaaatgctt gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct
     2221 catctgtaac atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat
     2281 cgggcttccc atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc
     2341 atttataccc atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg
     2401 tttcccgttg aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt
     2461 ttattgttca tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca
     2521 caacgtggct ttgttgaata aatcgaactt ttgctgagtt gaaggatcag ctcgagtgcc
     2581 acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac
     2641 gaggcagaat ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc
     2701 gcggccgctt ctagagacta gtggaagaca t
//
