LOCUS       sequence-136277-        7413 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Fluorescent reporter for transmembrane protein dimerization negative
            control..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7413)
  AUTHORS   Armstrong CR, Senes A
  TITLE     Screening for transmembrane association in divisome proteins using 
            TOXGREEN, a high-throughput variant of the TOXCAT assay.
  JOURNAL   Biochim Biophys Acta. 2016 Jul 21. pii: S0005-2736(16)30251-6. doi: 
            10.1016/j.bbamem.2016.07.008.
  PUBMED    27453198
REFERENCE   2  (bases 1 to 7413)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Biochim 
            Biophys Acta. 2016 Jul 21. pii: S0005-2736(16)30251-6. doi: 
            10.1016/j.bbamem.2016.07.008."
COMMENT     Sequence Label: pccGFPG83I
FEATURES             Location/Qualifiers
     source          1..7413
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(18..36)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     78..94
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     79..95
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             1014..2111
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1968..1977
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     2085..2108
                     /label=MBP-F
                     /note="Maltose binding protein, forward primer"
     promoter        2176..2280
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2281..2349,2350..3141)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2499..2518)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3312..3900
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3801..3820
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4054..4071
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4086..4226
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4152..4171)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4312..4334
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4328..4519)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     terminator      complement(5101..5128)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5220..5306)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      complement(5599..5626)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(5718..5804)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(6242..6955)
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MGKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      complement(7317..7403)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttcttgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
       61 aatggtttct tagacgttcg aggtcgacgg tatcgataag cttgatgggg atcgcgtttc
      121 tttattagtg gttgcagtct cgctcataat cgctccgttt acttctgttt caaacaattg
      181 atccattgag actcaatgga attaccttga tgtgcaagtg agatatggac aaaaaatgta
      241 aattcaaggt caaaactcat aaaaacactg ttttttgatc gagattggat tattctaagt
      301 ctgcattttt atcaaagaag ataaaaaaac cagtaaagtc tgagtgttgg gacagggaga
      361 tactgggaca ttagatgttc ggattaggac acaactcaaa agagatttcg atgagtcata
      421 ttggtactaa attcattctt gctgaaaaat ttaccttcga tcccctaagc aatactctga
      481 ttgacaaaga agatagtgaa gagatcattc gattaggcag caacgaaagc cgaattcttt
      541 ggctgctggc ccaacgtcca aacgaggtaa tttctcgcaa tgatttgcat gactttgttt
      601 ggcgagagca aggttttgaa gtcgatgatt ccagcttaac ccaagccatt tcgactctgc
      661 gcaaaatgct caaagattcg acaaagtccc cacaatacgt caaaacggtt ccgaagcgcg
      721 gttaccaatt gatcgcccga gtggaaacgg ttgaagaaga gatggctcgc gaaaacgaag
      781 ctgctcatga catctctcag ccagaatctg tcaatgaata cgcagaatca agcagtgtgc
      841 cttcatcagc cactgtagtg aacacaccgc agccagccaa tgtcgtggcg aataaatcgg
      901 ctccaaactt ggggaatcga gctagcctca ttatttttgg ggtgatggct attgttattg
      961 gaacgatcct gatcaaccca agccaatcca gctttaaacc cctaacggat atcaaaatcg
     1021 aagaaggtaa actggtaatc tggattaacg gcgataaagg ctataacggt ctcgctgaag
     1081 tcggtaagaa attcgagaaa gataccggaa ttaaagtcac cgttgagcat ccggataaac
     1141 tggaagagaa attcccacag gttgcggcaa ctggcgatgg ccctgacatt atcttctggg
     1201 cacacgaccg ctttggtggc tacgctcaat ctggcctgtt ggctgaaatc accccggaca
     1261 aagcgttcca ggacaagctg tatccgttta cctgggatgc cgtacgttac aacggcaagc
     1321 tgattgctta cccgatcgct gttgaagcgt tatcgctgat ttataacaaa gatctgctgc
     1381 cgaacccgcc aaaaacctgg gaagagatcc cggcgctgga taaagaactg aaagcgaaag
     1441 gtaagagcgc gctgatgttc aacctgcaag aaccgtactt cacctggccg ctgattgctg
     1501 ctgacggggg ttatgcgttc aagtatgaaa acggcaagta cgacattaaa gacgtgggcg
     1561 tggataacgc tggcgcgaaa gcgggtctga ccttcctggt tgacctgatt aaaaacaaac
     1621 acatgaatgc agacaccgat tactccatcg cagaagctgc ctttaataaa ggcgaaacag
     1681 cgatgaccat caacggcccg tgggcatggt ccaacatcga caccagcaaa gtgaattatg
     1741 gtgtaacggt actgccgacc ttcaagggtc aaccatccaa accgttcgtt ggcgtgctga
     1801 gcgcaggtat taacgccgcc agtccgaaca aagagctggc gaaagagttc ctcgaaaact
     1861 atctgctgac tgatgaaggt ctggaagcgg ttaataaaga caaaccgctg ggtgccgtag
     1921 cgctgaagtc ttacgaggaa gagttggcga aagatccacg tattgccgcc accatggaaa
     1981 acgcccagaa aggtgaaatc atgccgaaca tcccgcagat gtccgctttc tggtatgccg
     2041 tgcgtactgc ggtgatcaac gccgccagcg gtcgtcagac tgtcgatgaa gccctgaaag
     2101 acgcgcagac taattcgagc tcggtacccg gccggggatc ttaatctagc aggtggcact
     2161 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     2221 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt
     2281 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     2341 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     2401 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     2461 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     2521 cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     2581 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     2641 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     2701 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     2761 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     2821 cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     2881 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     2941 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
     3001 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
     3061 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     3121 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
     3181 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
     3241 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     3301 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     3361 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     3421 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     3481 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     3541 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     3601 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     3661 gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga aagcgccacg
     3721 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     3781 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     3841 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     3901 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     3961 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     4021 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     4081 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
     4141 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
     4201 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
     4261 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
     4321 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct
     4381 catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg tccagctcgt
     4441 tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg ttaagggcgg
     4501 ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc atgggggtaa
     4561 tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat gaacatgccc
     4621 ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa
     4681 aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta
     4741 gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct gacttccgcg
     4801 tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct caggtcgcag
     4861 acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca ttctgctaac
     4921 cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg atcatgcgca
     4981 cccgtggcca ggacccaacg ctgcccgact gctttcctga tgcaaaaacg aggctagttt
     5041 accgtatctg tggggggatg gcttgtagat atgacgacag gaagagtttg tagaaacgca
     5101 aaaaggccat ccgtcaggat ggccttctgc ttaatttgat gcctggcagt ttatggcggg
     5161 cgtcctgccc gccaccctcc gggccgttgc ttcgcaacgt tcaaatccgc tcccggcgga
     5221 tttgtcctac tcaggagagc gttcaccgac aaacaacaga taaaacgaaa ggcccagtct
     5281 ttcgactgag cctttcgttt tatttgatgc ctggcagttc cctactctcg catggggaga
     5341 ccccacacta ccatcggcgc tacggcgttt cacttctgag ttcggcatgg ggtcaggtgg
     5401 gaccaccgcg ctactgccgc caggcaaatt ctgttttatc agaccgcttc tgcgttctga
     5461 tttaatctgt atcaggctga aaatcttctc tcatccgcca aaacagctgc tttcctgatg
     5521 caaaaacgag gctagtttac cgtatctgtg gggggatggc ttgtagatat gacgacagga
     5581 agagtttgta gaaacgcaaa aaggccatcc gtcaggatgg ccttctgctt aatttgatgc
     5641 ctggcagttt atggcgggcg tcctgcccgc caccctccgg gccgttgctt cgcaacgttc
     5701 aaatccgctc ccggcggatt tgtcctactc aggagagcgt tcaccgacaa acaacagata
     5761 aaacgaaagg cccagtcttt cgactgagcc tttcgtttta tttgatgcct ggcagttccc
     5821 tactctcgca tggggagacc ccacactacc atcggcgcta cggcgtttca cttctgagtt
     5881 cggcatgggg tcaggtggga ccaccgcgct actgccgcca ggcaaattct gttttatcag
     5941 accgcttctg cgttctgatt taatctgtat caggctgaaa atcttctctc atccgccaaa
     6001 acagccgaga tgcgccgcgt gcggctgctg gagatggcgg acgcgatgga tatgttctgc
     6061 caagggttgg tttgcgcatt cacagttctc cgcaagaatt gattggctcc aattcttgga
     6121 gtggtgaatc cgttagcgag gtgccgcctc gacgaatttc tgccattcat ccgcttatta
     6181 tcacttattc aggcgtagca ccaggcgttt aagggcacca ataactgcct taaaaaaatc
     6241 atttgtacag ttcatccata ccatgcgtga tgcccgctgc ggttacgaac tccagcagaa
     6301 ccatgtgatc gcgtttctcg ttcggatctt tagacagaac gctttgcgtg ctcagatagt
     6361 gattgtctgg cagcagaaca ggaccatcac cgattggagt gttttgctgg tagtgatcag
     6421 ccagctgcac gctgccatcc tccacgttgt ggcgaatttt aaaattcgct ttaatgccat
     6481 ttttttgttt atcggcggtg atgtaaacat tgtggctgtt aaaattgtat tccagcttat
     6541 ggcccaggat attgccgtct tctttaaagt caatgccttt cagctcaatg cggtttacca
     6601 gggtatcgcc ttcaaatttc acttccgcac gcgttttgta cgtgccgtca tccttaaagg
     6661 aaatcgtgcg ttcctgcaca tagccttccg gcatggcgga cttgaagaag tcatgctgct
     6721 tcatgtggtc cggataacga gcaaagcact gaacaccata agtcagcgtc gttaccagag
     6781 tcggccaagg taccggcagt ttaccagtag tacagatgaa cttcagcgtc agtttaccat
     6841 tagttgcgtc accttcaccc tcgccacgca cggaaaactt atgaccgttg acatcaccat
     6901 ccagttccac cagaataggg acgacaccag tgaacagctc ttcgcctttg cccattttag
     6961 cttccttagc tcctgaaaat ctcgtcgaag ctcggcggat ttgtcctact caagcttgct
     7021 ccctttgttt aacagaaaaa taattgatca aaacaattaa ataagtaaat aaatttgtat
     7081 tatttgaaat caaaaatcaa aaatcaaaaa tcaaaaatca aaaatcaaaa atcaaaatat
     7141 actatttagt ccttttttat cattttctgt ttttatcggt aaaccccgtt tcacttctaa
     7201 gcttggctgc aggtcgacgg atcgatcccc gggaattccc tggcagttta tggcgggcgt
     7261 cctgcccgcc accctccggg ccgttgcttc gcaacgttca aatccgctcc cggcggattt
     7321 gtcctactca ggagagcgtt caccgacaaa caacagataa aacgaaaggc ccagtctttc
     7381 gactgagcct ttcgttttat ttgatgcctg gga
//
