LOCUS       sequence-174224-        4767 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GdT, a dimeric green fluorescent protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4767)
  AUTHORS   Wiens M, Shen Y, Li X, Salem M, Smisdom N, Zhang W, Brown A, 
            Campbell RE
  TITLE     A tandem green-red heterodimeric fluorescent protein with high FRET 
            efficiency.
  JOURNAL   Chembiochem. 2016 Oct 26. doi: 10.1002/cbic.201600492.
  PUBMED    27781394
REFERENCE   2  (bases 1 to 4767)
  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: 
            "Chembiochem. 2016 Oct 26. doi: 10.1002/cbic.201600492."
COMMENT     Sequence Label: pBad-HisB-GdT
FEATURES             Location/Qualifiers
     source          1..4767
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(53..931)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        958..1242
                     /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     1165..1184
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1288..1305
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1309..1341
                     /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             join(1345..1353,1354..1368)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             1384..2085
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPLLMYGSKMYVKHPADIPDYKKLSFPEGFKWVRVMNFEDGG
                     LVTATQDSSLQDGTLIYEVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTRVEQYERSEGRH
                     HLFLYDMDELYK"
     primer_bind     complement(1517..1535)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1785..1804
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1979..2002
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2125..2142)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2295..2381
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2473..2500
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2519..2610
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2611..2679,2680..3471)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2829..2848)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3642..4230
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4131..4150
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4384..4401
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4416..4556
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4482..4501)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4642..4664
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 tgcaaaccct atgctactcc gtcaagccgt caattgtctg attcgttacc aattatgaca
       61 acttgacggc tacatcattc actttttctt cacaaccggc acggaactcg ctcgggctgg
      121 ccccggtgca ttttttaaat acccgcgaga aatagagttg atcgtcaaaa ccaacattgc
      181 gaccgacggt ggcgataggc atccgggtgg tgctcaaaag cagcttcgcc tggctgatac
      241 gttggtcctc gcgccagctt aagacgctaa tccctaactg ctggcggaaa agatgtgaca
      301 gacgcgacgg cgacaagcaa acatgctgtg cgacgctggc gatatcaaaa ttgctgtctg
      361 ccaggtgatc gctgatgtac tgacaagcct cgcgtacccg attatccatc ggtggatgga
      421 gcgactcgtt aatcgcttcc atgcgccgca gtaacaattg ctcaagcaga tttatcgcca
      481 gcagctccga atagcgccct tccccttgcc cggcgttaat gatttgccca aacaggtcgc
      541 tgaaatgcgg ctggtgcgct tcatccgggc gaaagaaccc cgtattggca aatattgacg
      601 gccagttaag ccattcatgc cagtaggcgc gcggacgaaa gtaaacccac tggtgatacc
      661 attcgcgagc ctccggatga cgaccgtagt gatgaatctc tcctggcggg aacagcaaaa
      721 tatcacccgg tcggcaaaca aattctcgtc cctgattttt caccaccccc tgaccgcgaa
      781 tggtgagatt gagaatataa cctttcattc ccagcggtcg gtcgataaaa aaatcgagat
      841 aaccgttggc ctcaatcggc gttaaacccg ccaccagatg ggcattaaac gagtatcccg
      901 gcagcagggg atcattttgc gcttcagcca tacttttcat actcccgcca ttcagagaag
      961 aaaccaattg tccatattgc atcagacatt gccgtcactg cgtcttttac tggctcttct
     1021 cgctaaccaa accggtaacc ccgcttatta aaagcattct gtaacaaagc gggaccaaag
     1081 ccatgacaaa aacgcgtaac aaaagtgtct ataatcacgg cagaaaagtc cacattgatt
     1141 atttgcacgg cgtcacactt tgctatgcca tagcattttt atccataaga ttagcggatc
     1201 ctacctgacg ctttttatcg caactctcta ctgtttctcc atacccgttt tttgggctaa
     1261 caggaggaat taaccatggg gggttctcat catcatcatc atcatggtat ggctagcatg
     1321 actggtggac agcaaatggg tcgggatctg tacgacgatg acgataagga tccgagctcg
     1381 agaatggtga gcaagggcga ggaggtcatc aaagagttca tgcgcttcaa ggtgcgcatg
     1441 gagggctcca tgaacggcca cgagttcgag atcgagggcg agggcgaggg ccgcccctac
     1501 gagggcaccc agaccgccaa gctgaaggtg accaagggcg gccccctgcc cttcgcctgg
     1561 gacatcctgt cccccctgct catgtacggc tccaagatgt acgtgaagca ccccgccgac
     1621 atccccgatt acaagaagct gtccttcccc gagggcttca agtgggtgcg cgtgatgaac
     1681 ttcgaggacg gcggtctggt gaccgcgacc caggactcct ccctgcagga cggcacgctg
     1741 atctacgagg tgaagatgcg cggcaccaac ttcccccccg acggccccgt aatgcagaag
     1801 aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg cgtgctgaag
     1861 ggcgagatcc accaggccct gaagctgaag gacggcggcc actacctggt ggagttcaag
     1921 accatctaca tggccaagaa gcccgtgcaa ctgcccggct actactacgt ggacaccaag
     1981 ctggacatca cctcccacaa cgaggactac acccgtgtgg agcagtacga gcgctccgag
     2041 ggccgccacc acctgttcct gtacgacatg gacgagctgt acaagtaagc ttggctgttt
     2101 tggcggatga gagaagattt tcagcctgat acagattaaa tcagaacgca gaagcggtct
     2161 gataaaacag aatttgcctg gcggcagtag cgcggtggtc ccacctgacc ccatgccgaa
     2221 ctcagaagtg aaacgccgta gcgccgatgg tagtgtgggg tctccccatg cgagagtagg
     2281 gaactgccag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     2341 tctgttgttt gtcggtgaac gctctcctga gtaggacaaa tccgccggga gcggatttga
     2401 acgttgcgaa gcaacggccc ggagggtggc gggcaggacg cccgccataa actgccaggc
     2461 atcaaattaa gcagaaggcc atcctgacgg atggcctttt tgcgtttcta caaactcttt
     2521 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     2581 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     2641 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     2701 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     2761 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     2821 aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga gcaactcggt
     2881 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     2941 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     3001 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     3061 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     3121 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     3181 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     3241 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     3301 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     3361 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     3421 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     3481 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     3541 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     3601 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     3661 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     3721 gatcaagagc taccaactat ttttccgaag gtaactggct tcagcagagc gcagatacca
     3781 aatactgttc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     3841 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     3901 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     3961 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     4021 ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     4081 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     4141 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     4201 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     4261 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     4321 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     4381 cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt tctccttacg
     4441 catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg ctctgatgcc
     4501 gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg gctgcgcccc
     4561 gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatcagctt
     4621 acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac
     4681 cgaaacgcgc gaggcagcag atcaattcgc gcgcgaaggc gaagcggcat gcataatgtg
     4741 cctgtcaaat ggacgaagca gggattc
//
