LOCUS       sequence-250086-        3515 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the AAV 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 3515)
  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 3515)
  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_007
FEATURES             Location/Qualifiers
     source          1..3515
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             329..340
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             347..1060
                     /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      1072..1143
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1159..1186
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     repeat_region   1445..1585
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1445..1574
                     /label=AAV2 ITR
     CDS             complement(1625..2440)
                     /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(1625..2440)
                     /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     2348..2367
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2432..2451
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2665..3253
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3154..3173
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     repeat_region   3365..3505
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   3376..3505
                     /label=AAV2 ITR
ORIGIN
        1 gaattcgcat ctagactgat gagacgtggt agagccacaa acagccggta caagcaacga
       61 tctccaggac catctgaatc atgcgcggat gacacgaact cacgacggcg atcacagaca
      121 ttaacccaca gtacagacac tgcgacaacg tggcaattcg tcgcaataca acgtgagacc
      181 gaaagtgaaa cgtgatttca tgcgtcattt tgaacatttt gtaaatctta tttaataatg
      241 tgtgcggcaa ttcacattta atttatgaat gttttcttaa catcgcggca actcaagaaa
      301 cggcaggttc ggatcttagc tactagagaa agaggagaaa tactagatgc gtaaaggcga
      361 agagctgttc actggtgtcg tccctattct ggtggaactg gatggtgatg tcaacggtca
      421 taagttttcc gtgcgtggcg agggtgaagg tgacgcaact aatggtaaac tgacgctgaa
      481 gttcatctgt actactggta aactgccggt tccttggccg actctggtaa cgacgctgac
      541 ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg aagcagcatg acttcttcaa
      601 gtccgccatg ccggaaggct atgtgcagga acgcacgatt tcctttaagg atgacggcac
      661 gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc ctggtaaacc gcattgagct
      721 gaaaggcatt gactttaaag aggacggcaa tatcctgggc cataagctgg aatacaattt
      781 taacagccac aatgtttaca tcaccgccga taaacaaaaa aatggcatta aagcgaattt
      841 taaaattcgc cacaacgtgg aggatggcag cgtgcagctg gctgatcact accagcaaaa
      901 cactccaatc ggtgatggtc ctgttctgct gccagacaat cactatctga gcacgcaaag
      961 cgttctgtct aaagatccga acgagaaacg cgatcatatg gttctgctgg agttcgtaac
     1021 cgcagcgggc atcacgcatg gtatggatga actgtacaaa tgaccaggca tcaaataaaa
     1081 cgaaaggctc agtcgaaaga ctgggccttt cgttttatct gttgtttgtc ggtgaacgct
     1141 ctctactaga gtcacactgg ctcaccttcg ggtgggcctt tctgcgttta taggtctcag
     1201 ctggaaatct gctcgtcagt ggtgctcaca ctgacgaatc atgtacagat cataccgatg
     1261 actgcctggc gactcacaac taagcaagac agccggaacc agcgccggcg aacaccactg
     1321 catatatggc atatcacaac agtccacgtc tcaagcagtt acagagatgt tacgaaccac
     1381 tagtgcactg cagtacaact agagatctat gtgaggatcc taactcgaga tcgagagtgg
     1441 ccgcaggaac ccctagtgat ggagttggcc actccctctc tgcgcgctcg ctcgctcact
     1501 gaggccgggc gaccaaaggt cgcccgacgc ccgggctttg cccgggcggc ctcagtgagc
     1561 gagcgagcgc gcagctgcct gcaggccgag cggccgcgtg ttacaaccaa ttaaccaatt
     1621 ctgattagaa aaactcatcg agcatcaaat gaaactgcaa tttattcata tcaggattat
     1681 caataccata tttttgaaaa agccgtttct gtaatgaagg agaaaactca ccgaggcagt
     1741 tccataggat ggcaagatcc tggtatcggt ctgcgattcc gactcgtcca acatcaatac
     1801 aacctattaa tttcccctcg tcaaaaataa ggttatcaag tgagaaatca ccatgagtga
     1861 cgactgaatc cggtgagaat ggcaaaagct tatgcatttc tttccagact tgttcaacag
     1921 gccagccatt acgctcgtca tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg
     1981 attgcgcctg agcgaggcga aatacgcgat cgctgttaaa aggacaatta caaacaggaa
     2041 tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac aatattttca cctgaatcag
     2101 gatattcttc taatacctgg aatgctgttt tcccggggat cgcagtggtg agtaaccatg
     2161 catcatcagg agtacggata aaatgcttga tggtcggaag aggcataaat tccgtcagcc
     2221 agtttagtct gaccatctca tctgtaacat cattggcaac gctacctttg ccatgtttca
     2281 gaaacaactc tggcgcatcg ggcttcccat acaatcgata gattgtcgca cctgattgcc
     2341 cgacattatc gcgagcccat ttatacccat ataaatcagc atccatgttg gaatttaatc
     2401 gcggcctgga gcaagacgtt tcccgttgaa tatggctcat aacacccctt gtattactgt
     2461 ttatgtaagc agacagtttt attgttcatg atgatatatt tttatcttgt gcaatgtaac
     2521 atcagagatt ttgagacaca acgtggcttt gttgaataaa tcgaactttt gctgagttga
     2581 aggatcagtc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     2641 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     2701 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     2761 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     2821 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     2881 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     2941 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     3001 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     3061 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     3121 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     3181 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     3241 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     3301 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtgcggccgc
     3361 caatcctgca ggcagctgcg cgctcgctcg ctcactgagg ccgcccgggc aaagcccggg
     3421 cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag agagggagtg
     3481 gccaactcca tcactagggg ttcctgcggc cccct
//
