LOCUS       sequence-142928-        6140 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Localization: Protein Expression Vector , Excitation: 554, Emission:
            581.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6140)
  AUTHORS   Shaner NC, Campbell RE, Steinbach PA, Giepmans BN, Palmer AE, Tsien 
            RY
  TITLE     Improved monomeric red, orange and yellow fluorescent proteins 
            derived from Discosoma sp. red fluorescent protein.
  JOURNAL   Nat Biotechnol. 2004 Dec;22(12):1567-72. Epub 2004 Nov 21.
  PUBMED    15558047
REFERENCE   2  (bases 1 to 6140)
  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 
            Biotechnol."; date: "2004-12"; volume: "22(12)"; pages: "1567-72. 
            Epub 2004 Nov 21"
COMMENT     Sequence Label: tdTomato-pBAD
FEATURES             Location/Qualifiers
     source          1..6140
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     179..201
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     339..358
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(538..1416)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1443..1727
                     /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     1650..1669
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1754..1776
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1796..1813
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1817..1849
                     /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             1862..1882
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             1889..3319
                     /codon_start=1
                     /product="tandem dimeric (pseudo-monomeric) derivative of 
                     DsRed (Shaner et al., 2004)"
                     /label=tdTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLGHGTGSTGSGSSGTASSEDNNMAVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPY
                     EGTQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVM
                     NFEDGGLVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGV
                     LKGEIHQALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYE
                     RSEGRHHLFLYGMDELYK"
     primer_bind     complement(3365..3382)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3535..3621
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3713..3740
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3759..3850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3851..3919,3920..4711)
                     /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(4069..4088)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4753..5208
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4840..4859)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5050..5071
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5319..5907
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5808..5827
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6061..6078
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    join(6093..6140,1..93)
                     /label=bom
                     /note="basis of mobility region from pBR322"
ORIGIN
        1 catctggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
       61 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
      121 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
      181 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagaaggag
      241 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
      301 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
      361 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
      421 tgctcatgtt tgacagctta tcatcgatgc ataatgtgcc tgtcaaatgg acgaagcagg
      481 gattctgcaa accctatgct actccgtcaa gccgtcaatt gtctgattcg ttaccaatta
      541 tgacaacttg acggctacat cattcacttt ttcttcacaa ccggcacgga actcgctcgg
      601 gctggccccg gtgcattttt taaatacccg cgagaaatag agttgatcgt caaaaccaac
      661 attgcgaccg acggtggcga taggcatccg ggtggtgctc aaaagcagct tcgcctggct
      721 gatacgttgg tcctcgcgcc agcttaagac gctaatccct aactgctggc ggaaaagatg
      781 tgacagacgc gacggcgaca agcaaacatg ctgtgcgacg ctggcgatat caaaattgct
      841 gtctgccagg tgatcgctga tgtactgaca agcctcgcgt acccgattat ccatcggtgg
      901 atggagcgac tcgttaatcg cttccatgcg ccgcagtaac aattgctcaa gcagatttat
      961 cgccagcagc tccgaatagc gcccttcccc ttgcccggcg ttaatgattt gcccaaacag
     1021 gtcgctgaaa tgcggctggt gcgcttcatc cgggcgaaag aaccccgtat tggcaaatat
     1081 tgacggccag ttaagccatt catgccagta ggcgcgcgga cgaaagtaaa cccactggtg
     1141 ataccattcg cgagcctccg gatgacgacc gtagtgatga atctctcctg gcgggaacag
     1201 caaaatatca cccggtcggc aaacaaattc tcgtccctga tttttcacca ccccctgacc
     1261 gcgaatggtg agattgagaa tataaccttt cattcccagc ggtcggtcga taaaaaaatc
     1321 gagataaccg ttggcctcaa tcggcgttaa acccgccacc agatgggcat taaacgagta
     1381 tcccggcagc aggggatcat tttgcgcttc agccatactt ttcatactcc cgccattcag
     1441 agaagaaacc aattgtccat attgcatcag acattgccgt cactgcgtct tttactggct
     1501 cttctcgcta accaaaccgg taaccccgct tattaaaagc attctgtaac aaagcgggac
     1561 caaagccatg acaaaaacgc gtaacaaaag tgtctataat cacggcagaa aagtccacat
     1621 tgattatttg cacggcgtca cactttgcta tgccatagca tttttatcca taagattagc
     1681 ggatactacc tgacgctttt tatcgcaact ctctactgtt tctccatacc cgtttttttg
     1741 ggctagaaat aattttgttt aactttaaga aggagatata catatgcggg gttctcatca
     1801 tcatcatcat catggtatgg ctagcatgac tggtggacag caaatgggtc gggatctgta
     1861 cgagaacctg tacttccagg gctcgagcat ggtgagcaag ggcgaggagg tcatcaaaga
     1921 gttcatgcgc ttcaaggtgc gcatggaggg ctccatgaac ggccacgagt tcgagatcga
     1981 gggcgagggc gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa
     2041 gggcggcccc ctgcccttcg cctgggacat cctgtccccc cagttcatgt acggctccaa
     2101 ggcgtacgtg aagcaccccg ccgacatccc cgattacaag aagctgtcct tccccgaggg
     2161 cttcaagtgg gagcgcgtga tgaacttcga ggacggcggt ctggtgaccg tgacccagga
     2221 ctcctccctg caggacggca cgctgatcta caaggtgaag atgcgcggca ccaacttccc
     2281 ccccgacggc cccgtaatgc agaagaagac catgggctgg gaggcctcca ccgagcgcct
     2341 gtacccccgc gacggcgtgc tgaagggcga gatccaccag gccctgaagc tgaaggacgg
     2401 cggccactac ctggtggagt tcaagaccat ctacatggcc aagaagcccg tgcaactgcc
     2461 cggctactac tacgtggaca ccaagctgga catcacctcc cacaacgagg actacaccat
     2521 cgtggaacag tacgagcgct ccgagggccg ccaccacctg ttcctggggc atggcaccgg
     2581 cagcaccggc agcggcagct ccggcaccgc ctcctccgag gacaacaaca tggccgtcat
     2641 caaagagttc atgcgcttca aggtgcgcat ggagggctcc atgaacggcc acgagttcga
     2701 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     2761 gaccaagggc ggccccctgc ccttcgcctg ggacatcctg tccccccagt tcatgtacgg
     2821 ctccaaggcg tacgtgaagc accccgccga catccccgat tacaagaagc tgtccttccc
     2881 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggtctgg tgaccgtgac
     2941 ccaggactcc tccctgcagg acggcacgct gatctacaag gtgaagatgc gcggcaccaa
     3001 cttccccccc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctccaccga
     3061 gcgcctgtac ccccgcgacg gcgtgctgaa gggcgagatc caccaggccc tgaagctgaa
     3121 ggacggcggc cactacctgg tggagttcaa gaccatctac atggccaaga agcccgtgca
     3181 actgcccggc tactactacg tggacaccaa gctggacatc acctcccaca acgaggacta
     3241 caccatcgtg gaacagtacg agcgctccga gggccgccac cacctgttcc tgtacggcat
     3301 ggacgagctg tacaagtaag aattcgaagc ttggctgttt tggcggatga gagaagattt
     3361 tcagcctgat acagattaaa tcagaacgca gaagcggtct gataaaacag aatttgcctg
     3421 gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa ctcagaagtg aaacgccgta
     3481 gcgccgatgg tagtgtgggg tctccccatg cgagagtagg gaactgccag gcatcaaata
     3541 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
     3601 gctctcctga gtaggacaaa tccgccggga gcggatttga acgttgcgaa gcaacggccc
     3661 ggagggtggc gggcaggacg cccgccataa actgccaggc atcaaattaa gcagaaggcc
     3721 atcctgacgg atggcctttt tgcgtttcta caaactcttt tgtttatttt tctaaataca
     3781 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     3841 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3901 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3961 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     4021 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     4081 ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     4141 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     4201 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     4261 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     4321 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     4381 caccacgatg cctgcagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     4441 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     4501 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     4561 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     4621 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     4681 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     4741 ttagattgat ttacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     4801 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     4861 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     4921 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgatttg ggtgatggtt
     4981 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     5041 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcgggctatt
     5101 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     5161 aacaaaaatt taacgcgaat tttaacaaaa tattaacgtt tacaatttaa aaggatctag
     5221 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     5281 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     5341 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     5401 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     5461 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     5521 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     5581 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     5641 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     5701 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     5761 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     5821 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     5881 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     5941 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     6001 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     6061 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     6121 ctgtgcggta tttcacaccg
//
