LOCUS       sequence-429971-        3508 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  MoClo Destination Vector: (based on pSB1K3)  [A:LacZa:E].
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3508)
  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 3508)
  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_AE
FEATURES             Location/Qualifiers
     source          1..3508
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      21..78
                     /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(102..119)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(273..861)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(353..372)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(1094..1909)
                     /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(1094..1909)
                     /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     1817..1836
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1901..1920
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     mobile_element  1965..2732
                     /mobile_element_type="insertion sequence:IS1"
                     /label=IS1
                     /note="prokaryotic transposable element"
     primer_bind     2891..2909
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2913..2956)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2959..2980
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(2975..3298)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TSL"
     primer_bind     3193..3215
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3207..3224
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3208..3224
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    3225..3281
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(3294..3310)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3294..3310)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3307..3329)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3318..3334
                     /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(3342..3348,3349..3366,3367..3372))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3387..3408
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    join(3508,1..20)
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
ORIGIN
        1 actagtagcg gccgctgcag tccggcaaaa aagggcaagg tgtcaccacc ctgccctttt
       61 tctttaaaac cgaaaagatt acttcgcgtt atgcaggctt cctcgctcac tgactcgctg
      121 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
      181 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
      241 aggaaccgta aaaaggccgc gttgctggcg tttttccaca ggctccgccc ccctgacgag
      301 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
      361 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
      421 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
      481 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
      541 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
      601 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
      661 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
      721 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
      781 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
      841 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
      901 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
      961 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     1021 tggtctgaca gctcgagtcc cgtcaagtca gcgtaatgct ctgccagtgt tacaaccaat
     1081 taaccaattc tgattagaaa aactcatcga gcatcaaatg aaactgcaat ttattcatat
     1141 caggattatc aataccatat ttttgaaaaa gccgtttctg taatgaagga gaaaactcac
     1201 cgaggcagtt ccataggatg gcaagatcct ggtatcggtc tgcgattccg actcgtccaa
     1261 catcaataca acctattaat ttcccctcgt caaaaataag gttatcaagt gagaaatcac
     1321 catgagtgac gactgaatcc ggtgagaatg gcaaaagctt atgcatttct ttccagactt
     1381 gttcaacagg ccagccatta cgctcgtcat caaaatcact cgcatcaacc aaaccgttat
     1441 tcattcgtga ttgcgcctga gcgagacgaa atacgcgatc gctgttaaaa ggacaattac
     1501 aaacaggaat cgaatgcaac cggcgcagga acactgccag cgcatcaaca atattttcac
     1561 ctgaatcagg atattcttct aatacctgga atgctgtttt cccggggatc gcagtggtga
     1621 gtaaccatgc atcatcagga gtacggataa aatgcttgat ggtcggaaga ggcataaatt
     1681 ccgtcagcca gtttagtctg accatctcat ctgtaacatc attggcaacg ctacctttgc
     1741 catgtttcag aaacaactct ggcgcatcgg gcttcccata caatcgatag attgtcgcac
     1801 ctgattgccc gacattatcg cgagcccatt tatacccata taaatcagca tccatgttgg
     1861 aatttaatcg cggcctggag caagacgttt cccgttgaat atggctcata acaccccttg
     1921 tattactgtt tatgtaagca gacagtttta ttgttcatga tgatggtaat gactccaact
     1981 tattgatagt gttttatgtt cagataatgc ccgatgactt tgtcatgcag ctccaccgat
     2041 tttgagaacg acagcgactt ccgtcccagc cgtgccaggt gctgcctcag attcaggtta
     2101 tgccgctcaa ttcgctgcgt atatcgcttg ctgattacgt gcagctttcc cttcaggcgg
     2161 gattcataca gcggccagcc atccgtcatc catatcacca cgtcaaaggg tgacagcagg
     2221 ctcataagac gccccagcgt cgccatagtg cgttcaccga atacgtgcgc aacaaccgtc
     2281 ttccggagac tgtcatacgc gtaaaacagc cagcgctggc gcgatttagc cccgacatag
     2341 ccccactgtt cgtccatttc cgcgcagacg atgacgtcac tgcccggctg tatgcgcgag
     2401 gttaccgact gcggcctgag ttttttaagt gacgtaaaat cgtgttgagg ccaacgccca
     2461 taatgcgggc tgttgcccgg catccaacgc cattcatggc catatcaatg attttctggt
     2521 gcgtaccggg ttgagaagcg gtgtaagtga actgcagttg ccatgtttta cggcagtgag
     2581 agcagagata gcgctgatgt ccggcggtgc ttttgccgtt acgcaccacc ccgtcagtag
     2641 ctgaacagga gggacagctg atagaaacag aagccactgg agcacctcaa aaacaccatc
     2701 atacactaaa tcagtaagtt ggcagcatca cccatgatga tatattttta tcttgtgcaa
     2761 tgtaacatca gagattttga gacacaacgt ggctttgttg aataaatcga acttttgctg
     2821 agttgaagga tcagctcgag tgccacttga cgtctaagaa accattatta tcatgacatt
     2881 aacctataaa aataggcgta tcacgaggca gaatttcaga taaaaaaaat ccttagcttt
     2941 cgctaaggat gatttctgga attcgcggcc gcttctagag actagtggaa gacatggaga
     3001 gagacctgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     3061 tcaggcgcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3121 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     3181 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaattc gagctcggta
     3241 cccggggatc ctctagagtc gacctgcagg catgcaagct tggcgtaatc atggtcatag
     3301 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3361 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3421 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3481 cgcgcggggg tctctgctta tgtcttct
//
