LOCUS       sequence-250080-        4469 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the phiC31 attB GFP dropout destination vector as a type 0 
            part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4469)
  AUTHORS   Fonseca JP, Bonny AR, Kumar GR, Ng AH, Town J, Wu QC, Aslankoohi E, 
            Chen SY, Dods G, Harrigan P, Osimiri LC, Kistler AL, El-Samad H
  TITLE     A Toolkit for Rapid Modular Construction of Biological Circuits in 
            Mammalian Cells.
  JOURNAL   ACS Synth Biol. 2019 Nov 15;8(11):2593-2606. doi: 
            10.1021/acssynbio.9b00322. Epub 2019 Nov 5.
  PUBMED    31686495
REFERENCE   2  (bases 1 to 4469)
  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.9b00322"; journalName: "ACS Synth Biol"; date: 
            "2019-11-15- 15"; volume: "8"; issue: "11"; pages: "2593-2606"
COMMENT     Sequence Label: MTK0_013
FEATURES             Location/Qualifiers
     source          1..4469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     217..236
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    428..461
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     RBS             787..798
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             805..1518
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1530..1601
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1617..1644
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     protein_bind    2441..2474
                     /label=attB
                     /bound_moiety="phage phi-C31 integrase"
                     /note="minimal attB site for the phi-C31 integrase (Groth 
                     et al., 2000)"
     CDS             2479..2877
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     primer_bind     complement(2889..2906)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2895..3102
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(3157..3972)
                     /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(3157..3972)
                     /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     3880..3899
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3964..3983
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      join(4197..4469,1..316)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
       61 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
      121 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
      181 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
      241 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
      301 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
      361 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtgcggc
      421 cgccaatata acttcgtata atgtatgcta tacgaagtta tccctgaatt cgcatctaga
      481 ctgatgagac gtggtagagc cacaaacagc cggtacaagc aacgatctcc aggaccatct
      541 gaatcatgcg cggatgacac gaactcacga cggcgatcac agacattaac ccacagtaca
      601 gacactgcga caacgtggca attcgtcgca atacaacgga aagtgaaacg tgatttcatg
      661 cgtcattttg aacattttgt aaatcttatt taataatgtg tgcggcaatt cacatttaat
      721 ttatgaatgt tttcttaaca tcgcggcaac tcaagaaacg gcaggttcgg atcttagcta
      781 ctagagaaag aggagaaata ctagatgcgt aaaggcgaag agctgttcac tggtgtcgtc
      841 cctattctgg tggaactgga tggtgatgtc aacggtcata agttttccgt gcgtggcgag
      901 ggtgaaggtg acgcaactaa tggtaaactg acgctgaagt tcatctgtac tactggtaaa
      961 ctgccggttc cttggccgac tctggtaacg acgctgactt atggtgttca gtgctttgct
     1021 cgttatccgg accatatgaa gcagcatgac ttcttcaagt ccgccatgcc ggaaggctat
     1081 gtgcaggaac gcacgatttc ctttaaggat gacggcacgt acaaaacgcg tgcggaagtg
     1141 aaatttgaag gcgataccct ggtaaaccgc attgagctga aaggcattga ctttaaagag
     1201 gacggcaata tcctgggcca taagctggaa tacaatttta acagccacaa tgtttacatc
     1261 accgccgata aacaaaaaaa tggcattaaa gcgaatttta aaattcgcca caacgtggag
     1321 gatggcagcg tgcagctggc tgatcactac cagcaaaaca ctccaatcgg tgatggtcct
     1381 gttctgctgc cagacaatca ctatctgagc acgcaaagcg ttctgtctaa agatccgaac
     1441 gagaaacgcg atcatatggt tctgctggag ttcgtaaccg cagcgggcat cacgcatggt
     1501 atggatgaac tgtacaaatg accaggcatc aaataaaacg aaaggctcag tcgaaagact
     1561 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     1621 caccttcggg tgggcctttc tgcgtttata gctggaaatc tgctcgtcag tggtgctcac
     1681 actgacgaat catgtacaga tcataccgat gactgcctgg cgactcacaa ctaagcaaga
     1741 cagccggaac cagcgccggc gaacaccact gcatatatgg catatcacaa cagtccacgt
     1801 ctcaagcagt tacagagatg ttacgaacca ctagtgcact gcagatcgaa ttcgagttgg
     1861 cgcgcctgtc attctaaatc tctctttcag cctaaagctt tttccccgta tccccccagg
     1921 tgtctgcagg ctcaaagagc agcgagaagc gttcagagga aagcgatccc gtgccacctt
     1981 ccccgtgccc gggctgtccc cgcacgctgc cggctcgggg atgcgggggg agcgccggac
     2041 cggagcggag ccccgggcgg ctcgctgctg ccccctagcg ggggagggac gtaattacat
     2101 ccctgggggc tttgggaggg ggctgtcccc gtgagctccc aggcgcgcct gtcattctaa
     2161 atctctcttt cagcctaaag ctttttcccc gtatcccccc aggtgtctgc aggctcaaag
     2221 agcagcgaga agcgttcaga ggaaagcgat cccgtgccac cttccccgtg cccgggctgt
     2281 ccccgcacgc tgccggctcg gggatgcggg gggagcgccg gaccggagcg gagccccggg
     2341 cggctcgctg ctgcccccta gcgggggagg gacgtaatta catccctggg ggctttgggg
     2401 gggggctgtc cccgtgagct caataaaaga gcccactaca gtgccagggc gtgcccttgg
     2461 gctccccggg cgcggagtat ggccaagcct ttgtctcaag aagaatccac cctcattgaa
     2521 agagcaacgg ctacaatcaa cagcatcccc atctctgaag actacagcgt cgccagcgca
     2581 gctctctcta gcgacggccg catcttcact ggtgtcaatg tatatcattt tactggggga
     2641 ccttgtgcag aactcgtggt gctgggcact gctgctgctg cggcagctgg caacctgact
     2701 tgtatcgtcg cgatcggaaa tgagaacagg ggcatcttga gcccctgcgg acggtgccga
     2761 caggtgcttc tcgatctgca tcctgggatc aaagccatag tgaaggacag tgatggacag
     2821 ccgacggcag ttgggattcg tgaattgctg ccctctggtt atgtgtggga gggctaactc
     2881 gagcgacgcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     2941 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     3001 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     3061 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatccg
     3121 agcggccgcg tgttacaacc aattaaccaa ttctgattag aaaaactcat cgagcatcaa
     3181 atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa aaagccgttt
     3241 ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat cctggtatcg
     3301 gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct cgtcaaaaat
     3361 aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga atggcaaaag
     3421 cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt catcaaaatc
     3481 actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcgaggc gaaatacgcg
     3541 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
     3601 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
     3661 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
     3721 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
     3781 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
     3841 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
     3901 atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg
     3961 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
     4021 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
     4081 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag tcatgaccaa aatcccttaa
     4141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     4201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     4261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     4321 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     4381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4441 agtggcgata agtcgtgtct taccgggtt
//
