LOCUS       sequence-484672-        4236 bp    DNA     circular SYN 26-AUG-2025
DEFINITION  pBAD-mRFP Reporter Plasmid.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4236)
  TITLE     Phage-Assisted Evolution of Allosteric Protein Switches
REFERENCE   2  (bases 1 to 4236)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 26, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pJUMP27_pBAD_mRFP_LVA
FEATURES             Location/Qualifiers
     source          1..4236
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    47..66
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that start with ""ATG"""
     promoter        73..357
                     /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     280..299
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             397..1071
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     CDS             1072..1104
                     /codon_start=1
                     /product="C-terminal peptide that mediates degradation in 
                     bacteria through the ClpXP and ClpAP proteases (McGinness 
                     et al., 2006)"
                     /label=ssrA tag (LVA)
                     /note="mutant LVA variant that confers accelerated 
                     degradation under some conditions (Andersen et al., 1998)"
                     /translation="AANDENYALVA"
     terminator      1140..1211
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1227..1254
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      1330..1424
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1545..1564)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1556..2371
                     /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"
     oriT            2538..2646
                     /note="incP origin of transfer"
     CDS             complement(2657..3607)
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFDIFQWTNYAKFS
                     SEKLELVFSEEILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQLTSFEAKF
                     LSDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     rep_origin      3655..3877
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     terminator      complement(4144..4230)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
ORIGIN
        1 ttaattaagg agttttgcag gtgccttgga acacctgctt ttcgctgaat tcgcggccgc
       61 ttctagacta gaaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct
      121 tttactggct cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac
      181 aaagcgggac caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa
      241 aagtccacat tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca
      301 taagattagc ggatcctacc tgacgctttt tatcgcaact ctctactgtt tctccatgac
      361 cgaattcaaa agatctttta agaaggagat atacatatgg cgagtagcga agacgttatc
      421 aaagagttca tgcgtttcaa agttcgtatg gaaggttccg ttaacggtca cgagttcgaa
      481 atcgaaggtg aaggtgaagg tcgtccgtac gaaggtaccc agaccgctaa actgaaagtt
      541 accaaaggtg gtccgctgcc gttcgcttgg gacatcctgt ccccgcagtt ccagtacggt
      601 tccaaagctt acgttaaaca cccggctgac atcccggact acctgaaact gtccttcccg
      661 gaaggtttca aatgggaacg tgttatgaac ttcgaagacg gtggtgttgt taccgttacc
      721 caggactcct ccctgcaaga cggtgagttc atctacaaag ttaaactgcg tggtaccaac
      781 ttcccgtccg acggtccggt tatgcagaaa aaaaccatgg gttgggaagc ttccaccgaa
      841 cgtatgtacc cggaagacgg tgctctgaaa ggtgaaatca aaatgcgtct gaaactgaaa
      901 gacggtggtc actacgacgc tgaagttaaa accacctaca tggctaaaaa accggttcag
      961 ctgccgggtg cttacaaaac cgacatcaaa ctggacatca cctcccacaa cgaagactac
     1021 accatcgttg aacagtacga acgtgctgaa ggtcgtcact ccaccggtgc tgctgctaac
     1081 gacgaaaact acgctttagt agcttaagga tccaaactcg agtaaggatc tccaggcatc
     1141 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1201 tgaacgctct ctactagagt cacactggct caccttcggg tgggcctttc tgcgtttata
     1261 cctagggata tattccgctt cctcgctcac ctgcagggag ttgtcttcga agacttcgct
     1321 ctagtcttgg actcctgttg atagatccag taatgacctc agaactccat ctggatttgt
     1381 tcagaacgct cggttgccgc cgggcgtttt ttattggtga gaatccaggg gtccccaata
     1441 attacgattt aaatttgtgt ctcaaaatct ctgatgttac attgcacaag ataaaaatat
     1501 atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg gtgttatgag
     1561 ccatattcag cgtgaaacga gctgtagccg tccgcgtctg aacagcaaca tggatgcgga
     1621 tctgtatggc tataaatggg cgcgtgataa cgtgggtcag agcggcgcga ccatttatcg
     1681 tctgtatggc aaaccggatg cgccggaact gtttctgaaa catggcaaag gcagcgtggc
     1741 gaacgatgtg accgatgaaa tggtgcgtct gaactggctg accgaattta tgccgctgcc
     1801 gaccattaaa cattttattc gcaccccgga tgatgcgtgg ctgctgacca ccgcgattcc
     1861 gggcaaaacc gcgtttcagg tgctggaaga atatccggat agcggcgaaa acattgtgga
     1921 tgcgctggcc gtgtttctgc gtcgtctgca tagcattccg gtgtgcaact gcccgtttaa
     1981 cagcgatcgt gtgtttcgtc tggcccaggc gcagagccgt atgaacaacg gcctggtgga
     2041 tgcgagcgat tttgatgatg aacgtaacgg ctggccggtg gaacaggtgt ggaaagaaat
     2101 gcataaactg ctgccgttta gcccggatag cgtggtgacc cacggcgatt ttagcctgga
     2161 taacctgatt ttcgatgaag gcaaactgat tggctgcatt gatgtgggcc gtgtgggcat
     2221 tgcggatcgt tatcaggatc tggccattct gtggaactgc ctgggcgaat ttagcccgag
     2281 cctgcaaaaa cgtctgtttc agaaatatgg cattgataat ccggatatga acaaactgca
     2341 atttcatctg atgctggatg aatttttcta ataattaatt ggaccgcggt ccgcgcgttg
     2401 tccttttccg ctgcataacc ctgcttcggg gtcattatag cgattttttc ggtatatcca
     2461 tcctttttcg cacgatatac aggattttgc caaagggttc gtgtagactt tccttggtgt
     2521 atccaacggc gtcagccggg caggataggt gaagtaggcc cacccgcgag cgggtgttcc
     2581 ttcttcactg tcccttattc gcacctggcg gtgctcaacg ggaatcctgc tctgcgaggc
     2641 tggccgtagg ccggcctcag atccttccgt atttagccag tatgttctct agtgtggttc
     2701 gttgtttttg cgtgagccat gagaacgaac cattgagatc atacttactt tgcatgtcac
     2761 tcaaaaattt tgcctcaaaa ctggtgagct gaatttttgc agttaaagca tcgtgtagtg
     2821 tttttcttag tccgttatgt aggtaggaat ctgatgtaat ggttgttggt attttgtcac
     2881 cattcatttt tatctggttg ttctcaagtt cggttacgag atccatttgt ctatctagtt
     2941 caacttggaa aatcaacgta tcagtcgggc ggcctcgctt atcaaccacc aatttcatat
     3001 tgctgtaagt gtttaaatct ttacttattg gtttcaaaac ccattggtta agccttttaa
     3061 actcatggta gttattttca agcattaaca tgaacttaaa ttcatcaagg ctaatctcta
     3121 tatttgcctt gtgagttttc ttttgtgtta gttcttttaa taaccactca taaatcctca
     3181 tagagtattt gttttcaaaa gacttaacat gttccagatt atattttatg aattttttta
     3241 actggaaaag ataaggcaat atctcttcac taaaaactaa ttctaatttt tcgcttgaga
     3301 acttggcata gtttgtccac tggaaaatat caaagccttt aaccaaagga ttcctgattt
     3361 ccacagttct cgtcatcagc tctctggttg ctttagctaa tacaccataa gcattttccc
     3421 tactgatgtt catcatctga gcgtattggt tataagtgaa cgataccgtc cgttctttcc
     3481 ttgtagggtt ttcaatcgtg gggttgagta gtgccacaca gcataaaatt agcttggttt
     3541 catgctccgt taagtcatag cgactaatcg ctagttcatt tgctttgaaa acaactaatt
     3601 cagacataca tctcaattgg tctaggtgat tttaatcact ataccaattg agatgggcta
     3661 gtcaatgata attacatgtc cttttccttt gagttgtggg tatctgtaaa ttctgctaga
     3721 cctttgctgg aaaacttgta aattctgcta gaccctctgt aaattccgct agacctttgt
     3781 gtgttttttt tgtttatatt caagtggtta taatttatag aataaagaaa gaataaaaaa
     3841 agataaaaag aatagatccc agccctgtgt ataactcact actttagtca gttccgcagt
     3901 attacaaaag gatgtcgcaa acgctgtttg ctcctctaca aaacagacct taaaacccta
     3961 aaggcttaag tagcaccctc gcaagctcgg gcaaatcgct gaatattcct tttgtctccg
     4021 accatcaggc acctgagtcg ctgtcttttt cgtgacattc agttcgctgc gctcacggct
     4081 ctggcagtga atgggggtaa atggcactac aggcgcggcg cgcccagctg tctagggcgg
     4141 cggatttgtc ctactcagga gagcgttcac cgacaaacaa cagataaaac gaaaggccca
     4201 gtctttcgac tgagcctttc gttttatttg atgcct
//
