LOCUS pBad-HisB-GdT 4767 bp DNA circular SYN 26-AUG-2024
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 Aug 26, 2024 from SnapGene Server 7.0.3
https://www.biofeng.com
COMMENT SGRef: number: 1; type: "Journal Article"; journalName:
"Chembiochem. 2016 Oct 26. doi: 10.1002/cbic.201600492."
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 1345..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 2611..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
//
