LOCUS       sequence-271925-        2678 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the GFP dropout cassette for multi-TU destination vectors as
            a type 234 part to be used in the MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2678)
  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 2678)
  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: MTK234_008
FEATURES             Location/Qualifiers
     source          1..2678
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     469..488
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             831..842
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             849..1562
                     /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      1574..1645
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1661..1688
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(1712..1746)
                     /label=lambda t0 terminator
                     /note="minimal transcription terminator from phage lambda 
                     (Scholtissek and Grosse, 1987)"
     CDS             complement(1817..2476)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2391..2410
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      join(2658..2678,1..568)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
       61 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      121 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      181 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      241 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      301 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      361 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
      421 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
      481 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
      541 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
      601 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
      661 taaccgtagt cggtctcgaa cggaaagtga aacgtgattt catgcgtcat tttgaacatt
      721 ttgtaaatct tatttaataa tgtgtgcggc aattcacatt taatttatga atgttttctt
      781 aacatcgcgg caactcaaga aacggcaggt tcggatctta gctactagag aaagaggaga
      841 aatactagat gcgtaaaggc gaagagctgt tcactggtgt cgtccctatt ctggtggaac
      901 tggatggtga tgtcaacggt cataagtttt ccgtgcgtgg cgagggtgaa ggtgacgcaa
      961 ctaatggtaa actgacgctg aagttcatct gtactactgg taaactgccg gttccttggc
     1021 cgactctggt aacgacgctg acttatggtg ttcagtgctt tgctcgttat ccggaccata
     1081 tgaagcagca tgacttcttc aagtccgcca tgccggaagg ctatgtgcag gaacgcacga
     1141 tttcctttaa ggatgacggc acgtacaaaa cgcgtgcgga agtgaaattt gaaggcgata
     1201 ccctggtaaa ccgcattgag ctgaaaggca ttgactttaa agaggacggc aatatcctgg
     1261 gccataagct ggaatacaat tttaacagcc acaatgttta catcaccgcc gataaacaaa
     1321 aaaatggcat taaagcgaat tttaaaattc gccacaacgt ggaggatggc agcgtgcagc
     1381 tggctgatca ctaccagcaa aacactccaa tcggtgatgg tcctgttctg ctgccagaca
     1441 atcactatct gagcacgcaa agcgttctgt ctaaagatcc gaacgagaaa cgcgatcata
     1501 tggttctgct ggagttcgta accgcagcgg gcatcacgca tggtatggat gaactgtaca
     1561 aatgaccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     1621 ctgttgtttg tcggtgaacg ctctctacta gagtcacact ggctcacctt cgggtgggcc
     1681 tttctgcgtt tatagctggg agaccagacc aataaaaaac gcccggcggc aaccgagcgt
     1741 tctgaacaaa tccagatgga gttctgaggt cattactgga tctatcaaca ggagtccaag
     1801 cgagctcgat atcaaattac gccccgccct gccactcatc gcagtactgt tgtaattcat
     1861 taagcattct gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg
     1921 gcatcagcac cttgtcgcct tgcgtataat atttgcccat ggtgaaaacg ggggcgaaga
     1981 agttgtccat attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg
     2041 aaacgaaaaa catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac
     2101 acgccacatc ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc
     2161 agagcgatga aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat
     2221 cccatatcac cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca
     2281 ggcgggcaag aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct
     2341 ttaaaaaggc cgtaatatcc agctgaacgg tctggttata ggtacattga gcaactgact
     2401 gaaatgcctc aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag
     2461 tgattttttt ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata
     2521 cgcccggtag tgatcttatt tcattatggt gaaagttgga acctcttacg tgcccgatca
     2581 atcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2641 aagatcaaag gatcttcttg agatcctttt tttctgcg
//
