LOCUS       sequence-429967-        4069 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 4069)
  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 4069)
  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..4069
                     /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(2945..3760)
                     /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(2945..3760)
                     /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     3668..3687
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3752..3771
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     3965..3983
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(3987..4030)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    4033..4054
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(4049..4069,1..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 tctaaagtac tgagagatcc cctcataatt tccccaaaac gtaaccatgt
     1561 gtgaatagat tttgagtaag cagggttgca gccacgagtg agtcttccct tgttattgtg
     1621 tagccagaat gccgcaaaac ttccatgcct aagcgaactg ttgagagtac gtttcgattt
     1681 ctgactgtgt tagcctggaa gtgcttgtcc caaccttgtt tctgagcatg aacgcccgca
     1741 agccaacatg ttagttgaag catcagggcg attagcagca tgatatcaaa acgctctgag
     1801 ctgctcgttc ggctatggcg taggcctagt ccgtaggcag gacttttcaa gtctcggaag
     1861 gtttcttcaa tctgcattcg cttcgaatag atattaacaa gttgtttggg tgttcgaatt
     1921 tcaacaggta agttagttgc tagaatccat ggctcctttg ccgacgctga gtagatttta
     1981 ggtgacgggt ggtgacaatg agtccgtgtc gagcgctgat tttttcggcc tttagagcga
     2041 gatttataca atagaatttg gcatgagatt gggttgcttt tagtcagcct cttatagcct
     2101 aaagtctttg agtgactaga tgacatatca tgtaagttgc tgataggttt ccagttttcc
     2161 gctcctaggt ctgcatattg tacttttcct cttactcgac ttaaccagta ccaacccagc
     2221 ttctcaacgg atttatacca tggcacttta aagccagcat cactgacaat gagcggtgtg
     2281 gtgttactcg gtagaatgct cgcaaggtcg gctagaaatt ggtcatgagc tttctttgaa
     2341 cattgctctg aaagcgggaa cgctttctca taaagagtaa cagaacgacc gtgtagtgcg
     2401 actgaagctc gcaataccat aagccgtttt tgctcacgga tatcagacca gtcaacaagt
     2461 acaatgggca tcgtattgcc cgaacagata aagctagcat gccaacggta tacagcgagt
     2521 cgctctttgt ggaggtgacg attacctaac aatcggtcga ttcgtttgat gttatgtttt
     2581 gttctcgctt tggttggcag gttacggcca agttcggtaa gagtgagagt tttacagtca
     2641 agtaaggcgt ggcaagccaa cgttaagctg ttgagtcgtt ttaagtgtaa ttcggggcag
     2701 aattggtaaa gagagtcgtg taaaatatcg agttcgcaca ttttgttgtc tgattattga
     2761 tttttggcga aaccatttga tcatatgaca agatgtgtat ctaccttaac ttaatgattt
     2821 tgataaaaat cattagggga ttcatcagtc taaagtatat atgagtaaac ttggtctgac
     2881 agctcgagtc ccgtcaagtc agcgtaatgc tctgccagtg ttacaaccaa ttaaccaatt
     2941 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     3001 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     3061 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     3121 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     3181 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     3241 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     3301 attgcgcctg agcgagacga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     3361 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     3421 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     3481 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     3541 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     3601 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     3661 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     3721 gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     3781 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     3841 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     3901 aggatcagct cgagtgccac ttgacgtcta agaaaccatt attatcatga cattaaccta
     3961 taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag ctttcgctaa
     4021 ggatgatttc tggaattcgc ggccgcttct agagactagt ggaagacat
//
