LOCUS       sequence-250077-        8061 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the TU GFP dropout cassette with Ampicillin resistance and 
            bacterial artificial chromosome as a type 678 part to be used in the
            MTK system.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8061)
  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 8061)
  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: MTK678_002
FEATURES             Location/Qualifiers
     source          1..8061
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      96..684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     585..604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     RBS             933..944
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             951..1664
                     /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      1676..1747
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1763..1790
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     rep_origin      2433..2652
                     /label=ori2
                     /note="secondary origin of replication for the bacterial F 
                     plasmid; also known as oriS"
     CDS             2743..3498
                     /codon_start=1
                     /gene="repE"
                     /product="replication initiation protein for the bacterial 
                     F plasmid"
                     /label=repE
                     /translation="MAETAVINHKKRKNSPRIVQSNDLTEAAYSLSRDQKRMLYLFVDQ
                     IRKSDGTLQEHDGICEIHVAKYAEIFGLTSAEASKDIRQALKSFAGKEVVFYRPEEDAG
                     DEKGYESFPWFIKRAHSPSRGLYSVHINPYLIPFFIGLQNRFTQFRLSETKEITNPYAM
                     RLYESLCQYRKPDGSGIVSLKIDWIIERYQLPQSYQRMPDFRRRFLQVCVNEINSRTPM
                     RLSYIEKKKGRQTTHIVFSFRDITSMTTG"
     misc_feature    3501..3751
                     /gene="incC"
                     /label=incC
                     /note="incompatibility region of the bacterial F plasmid"
     CDS             4077..5252
                     /codon_start=1
                     /gene="sopA"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopA
                     /translation="MFRMKLMETLNQCINAGHEMTKAIAIAQFNDDSPEARKITRRWRI
                     GEAADLVGVSSQAIRDAEKAGRLPHPDMEIRGRVEQRVGYTIEQINHMRDVFGTRLRRA
                     EDVFPPVIGVAAHKGGVYKTSVSVHLAQDLALKGLRVLLVEGNDPQGTASMYHGWVPDL
                     HIHAEDTLLPFYLGEKDDVTYAIKPTCWPGLDIIPSCLALHRIETELMGKFDEGKLPTD
                     PHLMLRLAIETVAHDYDVIVIDSAPNLGIGTINVVCAADVLIVPTPAELFDYTSALQFF
                     DMLRDLLKNVDLKGFEPDVRILLTKYSNSNGSQSPWMEEQIRDAWGSMVLKNVVRETDE
                     VGKGQIRMRTVFEQAIDQRSSTGAWRNALSIWEPVCNEIFDRLIKPRWEIR"
     CDS             5252..6223
                     /codon_start=1
                     /gene="sopB"
                     /product="partitioning protein for the bacterial F plasmid"
                     /label=sopB
                     /translation="MKRAPVIPKHTLNTQPVEDTSLSTPAAPMVDSLIARVGVMARGNA
                     ITLPVCGRDVKFTLEVLRGDSVEKTSRVWSGNERDQELLTEDALDDLIPSFLLTGQQTP
                     AFGRRVSGVIEIADGSRRRKAAALTESDYRVLVGELDDEQMAALSRLGNDYRPTSAYER
                     GQRYASRLQNEFAGNISALADAENISRKIITRCINTAKLPKSVVALFSHPGELSARSGD
                     ALQKAFTDKEELLKQQASNLHEQKKAGVIFEAEEVITLLTSVLKTSSASRTSLSSRHQF
                     APGATVLYKGDKMVLNLDRSRVPTECIEKIEAILKELEKPAP"
     misc_feature    6296..6769
                     /gene="sopC"
                     /label=sopC
                     /note="centromere-like partitioning region of the bacterial
                     F plasmid"
     CDS             complement(join(7070..7861,7862..7930))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7693..7712
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7931..8035)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tgatacgccc cttgagacct catgaccaaa atcccttaac gtgagttttc gttccactga
       61 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
      121 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
      181 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
      241 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
      301 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
      361 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
      421 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
      481 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
      541 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
      601 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
      661 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
      721 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtgataacc
      781 gtatgaaagt gaaacgtgat ttcatgcgtc attttgaaca ttttgtaaat cttatttaat
      841 aatgtgtgcg gcaattcaca tttaatttat gaatgttttc ttaacatcgc ggcaactcaa
      901 gaaacggcag gttcggatct tagctactag agaaagagga gaaatactag atgcgtaaag
      961 gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg
     1021 gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc
     1081 tgaagttcat ctgtactact ggtaaactgc cggttccttg gccgactctg gtaacgacgc
     1141 tgacttatgg tgttcagtgc tttgctcgtt atccggacca tatgaagcag catgacttct
     1201 tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg
     1261 gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg
     1321 agctgaaagg cattgacttt aaagaggacg gcaatatcct gggccataag ctggaataca
     1381 attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga
     1441 attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc
     1501 aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc
     1561 aaagcgttct gtctaaagat ccgaacgaga aacgcgatca tatggttctg ctggagttcg
     1621 taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgacca ggcatcaaat
     1681 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1741 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataagtc
     1801 ggtctcatac ataccgctga aagttctgca aagcctgatg ggacataagt ccatcagttc
     1861 aacggaagtc tacacgaagg tttttgcgct ggatgtggct gcccggcacc gggtgcagtt
     1921 tgcgatgccg gagtctgatg cggttgcgat gctgaaacaa ttatcctgag aataaatgcc
     1981 ttggccttta tatggaaatg tggaactgag tggatatgct gtttttgtct gttaaacaga
     2041 gaagctggct gttatccact gagaagcgaa cgaaacagtc gggaaaatct cccattatcg
     2101 tagagatccg cattattaat ctcaggagcc tgtgtagcgt ttataggaag tagtgttctg
     2161 tcatgatgcc tgcaagcggt aacgaaaacg atttgaatat gccttcagga acaatagaaa
     2221 tcttcgtgcg gtgttacgtt gaagtggagc ggattatgtc agcaatggac agaacaacct
     2281 aatgaacaca gaaccatgat gtggtctgtc cttttacagc cagtagtgct cgccgcagtc
     2341 gagcgacagg gcgaagccct cgagtgagcg aggaagcacc agggaacagc acttatatat
     2401 tctgcttaca cacgatgcct gaaaaaactt cccttggggt tatccactta tccacgggga
     2461 tatttttata attatttttt ttatagtttt tagatcttct tttttagagc gccttgtagg
     2521 cctttatcca tgctggttct agagaaggtg ttgtgacaaa ttgccctttc agtgtgacaa
     2581 atcaccctca aatgacagtc ctgtctgtga caaattgccc ttaaccctgt gacaaattgc
     2641 cctcagaaga agctgttttt tcacaaagtt atccctgctt attgactctt ttttatttag
     2701 tgtgacaatc taaaaacttg tcacacttca catggatctg tcatggcgga aacagcggtt
     2761 atcaatcaca agaaacgtaa aaatagcccg cgaatcgtcc agtcaaacga cctcactgag
     2821 gcggcatata gtctctcccg ggatcaaaaa cgtatgctgt atctgttcgt tgaccagatc
     2881 agaaaatctg atggcaccct acaggaacat gacggtatct gcgagatcca tgttgctaaa
     2941 tatgctgaaa tattcggatt gacctctgcg gaagccagta aggatatacg gcaggcattg
     3001 aagagtttcg cggggaagga agtggttttt tatcgccctg aagaggatgc cggcgatgaa
     3061 aaaggctatg aatcttttcc ttggtttatc aaacgtgcgc acagtccatc cagagggctt
     3121 tacagtgtac atatcaaccc atatctcatt cccttcttta tcgggttaca gaaccggttt
     3181 acgcagtttc ggcttagtga aacaaaagaa atcaccaatc cgtatgccat gcgtttatac
     3241 gaatccctgt gtcagtatcg taagccggat ggctcaggca tcgtatctct gaaaatcgac
     3301 tggatcatag agcgttacca gctgcctcaa agttaccagc gtatgcctga cttccgccgc
     3361 cgcttcctgc aggtctgtgt taatgagatc aacagcagaa ctccaatgcg cctctcatac
     3421 attgagaaaa agaaaggccg ccagacgact catatcgtat tttccttccg cgatatcact
     3481 tccatgacga caggatagtc tgagggttat ctgtcacaga tttgagggtg gttcgtcaca
     3541 tttgttctga cctactgagg gtaatttgtc acagttttgc tgtttccttc agcctgcatg
     3601 gattttctca tactttttga actgtaattt ttaaggaagc caaatttgag ggcagtttgt
     3661 cacagttgat ttccttctct ttcccttcgt catgtgacct gatatcgggg gttagttcgt
     3721 catcattgat gagggttgat tatcacagtt tattactctg aattggctat ccgcgtgtgt
     3781 acctctacct ggagtttttc ccacggtgga tatttcttct tgcgctgagc gtaagagcta
     3841 tctgacagaa cagttcttct ttgcttcctc gccagttcgc tcgctatgct cggttacacg
     3901 gctgcggcga gcgctagtga taataagtga ctgaggtatg tgctcttctt atctcctttt
     3961 gtagtgttgc tcttatttta aacaactttg cggttttttg atgactttgc gattttgttg
     4021 ttgctttgca gtaaattgca agatttaata aaaaaacgca aagcaatgat taaaggatgt
     4081 tcagaatgaa actcatggaa acacttaacc agtgcataaa cgctggtcat gaaatgacga
     4141 aggctatcgc cattgcacag tttaatgatg acagcccgga agcgaggaaa ataacccggc
     4201 gctggagaat aggtgaagca gcggatttag ttggggtttc ttctcaggct atcagagatg
     4261 ccgagaaagc agggcgacta ccgcacccgg atatggaaat tcgaggacgg gttgagcaac
     4321 gtgttggtta tacaattgaa caaattaatc atatgcgtga tgtgtttggt acgcgattgc
     4381 gacgtgctga agacgtattt ccaccggtga tcggggttgc tgcccataaa ggtggcgttt
     4441 acaaaacctc agtttctgtt catcttgctc aggatctggc tctgaagggg ctacgtgttt
     4501 tgctcgtgga aggtaacgac ccccagggaa cagcctcaat gtatcacgga tgggtaccag
     4561 atcttcatat tcatgcagaa gacactctcc tgcctttcta tcttggggaa aaggacgatg
     4621 tcacttatgc aataaagccc acttgctggc cggggcttga cattattcct tcctgtctgg
     4681 ctctgcaccg tattgaaact gagttaatgg gcaaatttga tgaaggtaaa ctgcccaccg
     4741 atccacacct gatgctccga ctggccattg aaactgttgc tcatgactat gatgtcatag
     4801 ttattgacag cgcgcctaac ctgggtatcg gcacgattaa tgtcgtatgt gctgctgatg
     4861 tgctgattgt tcccacgcct gctgagttgt ttgactacac ctccgcactg cagtttttcg
     4921 atatgcttcg tgatctgctc aagaacgttg atcttaaagg gttcgagcct gatgtacgta
     4981 ttttgcttac caaatacagc aatagtaatg gctctcagtc cccgtggatg gaggagcaaa
     5041 ttcgggatgc ctggggaagc atggttctaa aaaatgttgt acgtgaaacg gatgaagttg
     5101 gtaaaggtca gatccggatg agaactgttt ttgaacaggc cattgatcaa cgctcttcaa
     5161 ctggtgcctg gagaaatgct ctttctattt gggaacctgt ctgcaatgaa attttcgatc
     5221 gtctgattaa accacgctgg gagattagat aatgaagcgt gcgcctgtta ttccaaaaca
     5281 tacgctcaat actcaaccgg ttgaagatac ttcgttatcg acaccagctg ccccgatggt
     5341 ggattcgtta attgcgcgcg taggagtaat ggctcgcggt aatgccatta ctttgcctgt
     5401 atgtggtcgg gatgtgaagt ttactcttga agtgctccgg ggtgatagtg ttgagaagac
     5461 ctctcgggta tggtcaggta atgaacgtga ccaggagctg cttactgagg acgcactgga
     5521 tgatctcatc ccttcttttc tactgactgg tcaacagaca ccggcgttcg gtcgaagagt
     5581 atctggtgtc atagaaattg ccgatgggag tcgccgtcgt aaagctgctg cacttaccga
     5641 aagtgattat cgtgttctgg ttggcgagct ggatgatgag cagatggctg cattatccag
     5701 attgggtaac gattatcgcc caacaagtgc ttatgaacgt ggtcagcgtt atgcaagccg
     5761 attgcagaat gaatttgctg gaaatatttc tgcgctggct gatgcggaaa atatttcacg
     5821 taagattatt acccgctgta tcaacaccgc caaattgcct aaatcagttg ttgctctttt
     5881 ttctcacccc ggtgaactat ctgcccggtc aggtgatgca cttcaaaaag cctttacaga
     5941 taaagaggaa ttacttaagc agcaggcatc taaccttcat gagcagaaaa aagctggggt
     6001 gatatttgaa gctgaagaag ttatcactct tttaacttct gtgcttaaaa cgtcatctgc
     6061 atcaagaact agtttaagct cacgacatca gtttgctcct ggagcgacag tattgtataa
     6121 gggcgataaa atggtgctta acctggacag gtcacgtgtt ccaactgagt gtatagagaa
     6181 aattgaggcc attcttaagg aacttgaaaa gccagcaccc tgatgcgacc acgttttagt
     6241 ctacgtttat ctgtctttac ttaatgtcct ttgttacagg ccagaaagca taactggcct
     6301 gaatattctc tctgggccca ctgttccact tgtatcgtcg gtctgataat cagactggga
     6361 ccacggtccc actcgtatcg tcggtctgat tattagtctg ggaccacggt cccactcgta
     6421 tcgtcggtct gattattagt ctgggaccac ggtcccactc gtatcgtcgg tctgataatc
     6481 agactgggac cacggtccca ctcgtatcgt cggtctgatt attagtctgg gaccatggtc
     6541 ccactcgtat cgtcggtctg attattagtc tgggaccacg gtcccactcg tatcgtcggt
     6601 ctgattatta gtctggaacc acggtcccac tcgtatcgtc ggtctgatta ttagtctggg
     6661 accacggtcc cactcgtatc gtcggtctga ttattagtct gggaccacga tcccactcgt
     6721 gttgtcggtc tgattatcgg tctgggacca cggtcccact tgtattgtcg atcagactat
     6781 cagcgtgaga ctacgattcc atcaatgcct gtcaagggca agtattgaca tgtcgtcgta
     6841 acctgtagaa cggagtaacc tcggtgtgcg gttgtatgcc tgctgtggat tgctgctgtg
     6901 tcctgcttat ccacaacatt ttgcgcacgg ttatgtggac aaaatacctg gttacccagg
     6961 ccgtgccggc acgttgatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7021 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7081 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7141 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7201 tgataccgcg ggacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7261 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7321 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7381 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7441 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7501 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7561 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7621 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7681 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7741 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7801 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     7861 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     7921 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     7981 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     8041 ttccccgaaa agtgccacct g
//
