LOCUS       sequence-67701-e        4939 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Contains PBAD::GFP reporter, used for heterodimer competition assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4939)
  AUTHORS   Su PC, Berger BW
  TITLE     A Novel Assay for Assessing Juxtamembrane and Transmembrane Domain 
            Interactions Important for Receptor Heterodimerization.
  JOURNAL   J Mol Biol. 2013 Jul 19. pii: S0022-2836(13)00456-7. doi: 
            10.1016/j.jmb.2013.07.022.
  PUBMED    23876708
REFERENCE   2  (bases 1 to 4939)
  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"; journalName: "J Mol Biol.
            2013 Jul 19. pii: S0022-2836(13)00456-7. doi: 
            10.1016/j.jmb.2013.07.022."
COMMENT     Sequence Label: pAraGFPCDF
FEATURES             Location/Qualifiers
     source          1..4939
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        221..505
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     428..447
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             566..1279
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFAYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     1219..1239
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        1372..1390
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1391..1415
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     CDS             1524..1568
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(1606..1624)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1620..1667
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(1838..2629)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     promoter        complement(2630..2721)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2769..3507)
                     /direction=LEFT
                     /label=CloDF13 ori
                     /note="Plasmids containing the CloDF13 (CDF) origin of 
                     replication can be propagated in E. coli cells that contain
                     additional plasmids with compatible origins."
     CDS             complement(3717..4799)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=lacI
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4761..4780
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4800..4877)
                     /gene="lacI"
                     /label=lacI promoter
     promoter        join(4923..4939,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct gtagaaataa ttttgtttaa ctttaataag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgaattcatc
      121 ggcgttaaac ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt
      181 tgcgcttcag ccatactttt catactcccg ccattcagag aagaaaccaa ttgtccatat
      241 tgcatcagac attgccgtca ctgcgtcttt tactggctct tctcgctaac caaaccggta
      301 accccgctta ttaaaagcat tctgtaacaa agcgggacca aagccatgac aaaaacgcgt
      361 aacaaaagtg tctataatca cggcagaaaa gtccacattg attatttgca cggcgtcaca
      421 ctttgctatg ccatagcatt tttatccata agattagcgg atcctacctg acgcttttta
      481 tcgcaactct ctactgtttc tccatacccg tttttttggg aaaaaaaccc ggggatcctc
      541 tagatttaat gaaggagata tacatatgag taaaggagaa gaacttttca ctggagtggt
      601 cccaattctt gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga
      661 gggtgaaggt gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa
      721 actacctgtt ccatggccaa cacttgtcac tactttcgcg tatggtcttc aatgctttgc
      781 gagataccca gatcatatga aacagcatga ctttttcaag agtgccatgc ccgaaggtta
      841 tgtacaggaa agaactatat ttttcaaaga tgacgggaac tacaagacac gtgctgaagt
      901 caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg attttaaaga
      961 agatggaaac attcttggac acaaattgga atacaactat aactcacaca atgtatacat
     1021 catggcagac aaacaaaaga atggaatcaa agttaacttc aaaattagac acaacattaa
     1081 agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg gcgatggccc
     1141 tgtcctttta ccagacaacc attacctgtc cacacaatct gccctttcga aagatcccaa
     1201 cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga ttacacatgg
     1261 catggatgaa ctatacaaat aattttttct gcaggtcgac aagcttgcgg ccgcataatg
     1321 cttaagtcga acagaaagta atcgtattgt acacggccgc ataatcgaaa ttaatacgac
     1381 tcactatagg ggaattgtga gcggataaca attccccatc ttagtatatt agttaagtat
     1441 aagaaggaga tatacatatg gcagatctca attggatatc ggccggccac gcgatcgctg
     1501 acgtcggtac cctcgagtct ggtaaagaaa ccgctgctgc gaaatttgaa cgccagcaca
     1561 tggactcgtc tactagcgca gcttaattaa cctaggctgc tgccaccgct gagcaataac
     1621 tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaacctcagg
     1681 catttgagaa gcacacggtc acactgcttc cggtagtcaa taaaccggta aaccagcaat
     1741 agacataagc ggctatttaa cgaccctgcc ctgaaccgac gaccgggtca tcgtggccgg
     1801 atcttgcggc ccctcggctt gaacgaattg ttagacatta tttgccgact accttggtga
     1861 tctcgccttt cacgtagtgg acaaattctt ccaactgatc tgcgcgcgag gccaagcgat
     1921 cttcttcttg tccaagataa gcctgtctag cttcaagtat gacgggctga tactgggccg
     1981 gcaggcgctc cattgcccag tcggcagcga catccttcgg cgcgattttg ccggttactg
     2041 cgctgtacca aatgcgggac aacgtaagca ctacatttcg ctcatcgcca gcccagtcgg
     2101 gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc aaatagatcc tgttcaggaa
     2161 ccggatcaaa gagttcctcc gccgctggac ctaccaaggc aacgctatgt tctcttgctt
     2221 ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg ctcgaagata cctgcaagaa
     2281 tgtcattgcg ctgccattct ccaaattgca gttcgcgctt agctggataa cgccacggaa
     2341 tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg gagaatctcg ctctctccag
     2401 gggaagccga agtttccaaa aggtcgttga tcaaagctcg ccgcgttgtt tcatcaagcc
     2461 ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg cttcaggccg ccatccactg
     2521 cggagccgta caaatgtacg gccagcaacg tcggttcgag atggcgctcg atgacgccaa
     2581 ctacctctga tagttgagtc gatacttcgg cgatcaccgc ttccctcata ctcttccttt
     2641 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2701 gtatttagaa aaataaacaa atagctagct cactcggtcg ctacgctccg ggcgtgagac
     2761 tgcggcgggc gctgcggaca catacaaagt tacccacaga ttccgtggat aagcagggga
     2821 ctaacatgtg aggcaaaaca gcagggccgc gccggtggcg tttttccata ggctccgccc
     2881 tcctgccaga gttcacataa acagacgctt ttccggtgca tctgtgggag ccgtgaggct
     2941 caaccatgaa tctgacagta cgggcgaaac ccgacaggac ttaaagatcc ccaccgtttc
     3001 cggcgggtcg ctccctcttg cgctctcctg ttccgaccct gccgtttacc ggatacctgt
     3061 tccgcctttc tcccttacgg gaagtgtggc gctttctcat agctcacaca ctggtatctc
     3121 ggctcggtgt aggtcgttcg ctccaagctg ggctgtaagc aagaactccc cgttcagccc
     3181 gactgctgcg ccttatccgg taactgttca cttgagtcca acccggaaaa gcacggtaaa
     3241 acgccactgg cagcagccat tggtaactgg gagttcgcag aggatttgtt tagctaaaca
     3301 cgcggttgct cttgaagtgt gcgccaaagt ccggctacac tggaaggaca gatttggttg
     3361 ctgtgctctg cgaaagccag ttaccacggt taagcagttc cccaactgac ttaaccttcg
     3421 atcaaaccac ctccccaggt ggttttttcg tttacagggc aaaagattac gcgcagaaaa
     3481 aaaggatctc aagaagatcc tttgatcttt tctactgaac cgctctagat ttcagtgcaa
     3541 tttatctctt caaatgtagc acctgaagtc agccccatac gatataagtt gtaattctca
     3601 tgttagtcat gccccgcgcc caccggaagg agctgactgg gttgaaggct ctcaagggca
     3661 tcggtcgaga tcccggtgcc taatgagtga gctaacttac attaattgcg ttgcgctcac
     3721 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     3781 cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac cagtgagacg
     3841 ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa gcggtccacg
     3901 ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg gatataacat
     3961 gagctgtctt cggtatcgtc gtatcccact accgagatgt ccgcaccaac gcgcagcccg
     4021 gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac cagcatcgca
     4081 gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga catggcactc
     4141 cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata tttatgccag
     4201 ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag cgcgatttgc
     4261 tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc atgggagaaa
     4321 ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg aacattagtg
     4381 caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat gatcagccca
     4441 ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac gccgcttcgt
     4501 tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt aatcgccgcg
     4561 acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat cagcaacgac
     4621 tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc
     4681 gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac cacgcgggaa
     4741 acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac tggtttcaca
     4801 ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg aaaggttttg
     4861 cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct gcattaggaa
     4921 attaatacga ctcactata
//
