LOCUS       sequence-458428-        5423 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 559, Emission:
            600.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5423)
  AUTHORS   Lam AJ, St-Pierre F, Gong Y, Marshall JD, Cranfill PJ, Baird MA, 
            McKeown MR, Wiedenmann J, Davidson MW, Schnitzer MJ, Tsien RY, Lin 
            MZ
  TITLE     Improving FRET dynamic range with bright green and red fluorescent 
            proteins.
  JOURNAL   Nat Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171.
  PUBMED    22961245
REFERENCE   2  (bases 1 to 5423)
  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: "Nat 
            Methods. 2012 Sep 9. doi: 10.1038/nmeth.2171."
COMMENT     Sequence Label: mRuby2-pBAD
FEATURES             Location/Qualifiers
     source          1..5423
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             312..334
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             354..371
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             375..407
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             420..440
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             447..1157
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     primer_bind     complement(1206..1223)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1376..1462
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1554..1581
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1600..1691
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1692..1760,1761..2552)
                     /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(1910..1929)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2594..3049
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2681..2700)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2891..2912
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      3160..3748
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3649..3668
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3902..3919
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3934..4074
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4000..4019)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4160..4182
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4320..4339
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4519..5397)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atactacctg acgcttttta tcgcaactct ctactgtttc tccatacccg tttttttggg
      301 ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc
      361 atcatcatca tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg
      421 agaacctgta cttccagggc tcgagcatgg tgtctaaggg cgaagagctg atcaaggaaa
      481 atatgcgtat gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag
      541 gtgaaggaga aggcaatccg tacatgggaa ctcaaaccat gaggatcaaa gtcatcgagg
      601 gaggacccct gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta
      661 cttttatcaa gtacccgaaa ggcattcctg atttctttaa acagtccttt cctgagggtt
      721 ttacttggga aagagttacg agatacgaag atggtggagt cgtcaccgtc atgcaggaca
      781 ccagccttga ggatggctgt ctcgtttacc acgtccaagt cagaggggta aactttccct
      841 ccaatggtcc cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt
      901 atccagcaga tggtggtctg aggggataca ctcatatggc actgaaagtt gatggtggtg
      961 gccatctgtc ttgctctttc gtaacaactt acaggtcaaa aaagaccgtc gggaacatca
     1021 agatgcccgg tatccatgcc gttgatcacc gcctggaaag gttagaggaa agtgacaatg
     1081 aaatgttcgt agtacaacgc gaacacgcag ttgccaagtt cgccgggctt ggtggtggga
     1141 tggacgagct gtacaagtaa gaattcgaag cttggctgtt ttggcggatg agagaagatt
     1201 ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca gaatttgcct
     1261 ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt gaaacgccgt
     1321 agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca ggcatcaaat
     1381 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1441 cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga agcaacggcc
     1501 cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta agcagaaggc
     1561 catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt ttctaaatac
     1621 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     1681 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
     1741 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
     1801 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
     1861 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1921 cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1981 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     2041 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     2101 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     2161 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     2221 acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     2281 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     2341 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     2401 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     2461 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     2521 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     2581 tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2641 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     2701 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     2761 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     2821 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     2881 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     2941 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3001 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta aaaggatcta
     3061 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     3121 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     3181 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     3241 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     3301 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     3361 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     3421 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     3481 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     3541 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     3601 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     3661 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     3721 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     3781 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     3841 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     3901 cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca
     3961 tctgtgcggt atttcacacc gcatctggtg cactctcagt acaatctgct ctgatgccgc
     4021 atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga
     4081 cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac
     4141 agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg
     4201 aaacgcgcga ggcagaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     4261 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     4321 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     4381 atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg cataatgtgc
     4441 ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca agccgtcaat
     4501 tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt tttcttcaca
     4561 accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc gcgagaaata
     4621 gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc gggtggtgct
     4681 caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga cgctaatccc
     4741 taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat gctgtgcgac
     4801 gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac aagcctcgcg
     4861 tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc gccgcagtaa
     4921 caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc cttgcccggc
     4981 gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat ccgggcgaaa
     5041 gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt aggcgcgcgg
     5101 acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac cgtagtgatg
     5161 aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt ctcgtccctg
     5221 atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt tcattcccag
     5281 cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta aacccgccac
     5341 cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt cagccatact
     5401 tttcatactc ccgccattca gag
//
