LOCUS       sequence-174222-        5481 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses GRvT, a tandem heterodimer green and red fluorescent 
            protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5481)
  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 5481)
  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-GRvT
FEATURES             Location/Qualifiers
     source          1..5481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(47..925)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQEVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        952..1236
                     /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     1159..1178
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             1282..1299
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1303..1335
                     /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(1339..1347,1348..1362)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1511..1529)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1779..1798
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1973..1996
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2833..2850)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3003..3089
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3181..3208
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3227..3318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3319..3387,3388..4179)
                     /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(3537..3556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4350..4938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4839..4858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5092..5109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5124..5264
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5190..5209)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5350..5372
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 ccctatgcta ctccgtcaag ccgtcaattg tctgattcgt taccaattat gacaacttga
       61 cggctacatc attcactttt tcttcacaac cggcacggaa ctcgctcggg ctggccccgg
      121 tgcatttttt aaatacccgc gagaaataga gttgatcgtc aaaaccaaca ttgcgaccga
      181 cggtggcgat aggcatccgg gtggtgctca aaagcagctt cgcctggctg atacgttggt
      241 cctcgcgcca gcttaagacg ctaatcccta actgctggcg gaaaagatgt gacagacgcg
      301 acggcgacaa gcaaacatgc tgtgcgacgc tggcgatatc aaaattgctg tctgccaggt
      361 gatcgctgat gtactgacaa gcctcgcgta cccgattatc catcggtgga tggagcgact
      421 cgttaatcgc ttccatgcgc cgcagtaaca attgctcaag cagatttatc gccagcagct
      481 ccgaatagcg cccttcccct tgcccggcgt taatgatttg cccaaacagg tcgctgaaat
      541 gcggctggtg cgcttcatcc gggcgaaaga accccgtatt ggcaaatatt gacggccagt
      601 taagccattc atgccagtag gcgcgcggac gaaagtaaac ccactggtga taccattcgc
      661 gagcctccgg atgacgaccg tagtgatgaa tctctcctgg cgggaacagc aaaatatcac
      721 ccggtcggca aacaaattct cgtccctgat ttttcaccac ctcctgaccg cgaatggtga
      781 gattgagaat ataacctttc attcccagcg gtcggtcgat aaaaaaatcg agataaccgt
      841 tggcctcaat cggcgttaaa cccgccacca gatgggcatt aaacgagtat cccggcagca
      901 ggggatcatt ttgcgcttca gccatacttt tcatactccc gccattcaga gaagaaacca
      961 attgtccata ttgcatcaga cattgccgtc actgcgtctt ttactggctc ttctcgctaa
     1021 ccaaaccggt aaccccgctt attaaaagca ttctgtaaca aagcgggacc aaagccatga
     1081 caaaaacgcg taacaaaagt gtctataatc acggcagaaa agtccacatt gattatttgc
     1141 acggcgtcac actttgctat gccatagcat ttttatccat aagattagcg gatcctacct
     1201 gacgcttttt atcgcaactc tctactgttt ctccataccc gttttttggg ctaacaggag
     1261 gaattaacca tggggggttc tcatcatcat catcatcatg gtatggctag catgactggt
     1321 ggacagcaaa tgggtcggga tctgtacgac gatgacgata aggatccgag ctcgagaatg
     1381 gtgagcaagg gcgaggaggt catcaaagag ttcatgcgct tcaaggtgcg catggagggc
     1441 tccatgaacg gccacgagtt cgagatcgag ggcgagggcg agggccgccc ctacgagggc
     1501 acccagaccg ccaagctgaa ggtgaccaag ggcggccccc tgcccttcgc ctgggacatc
     1561 ctgtcccccc tgctcatgta cggctccaag atgtacgtga agcaccccgc cgacatcccc
     1621 gattacaaga agctgtcctt ccccgagggc ttcaagtggg tgcgcgtgat gaacttcgag
     1681 gacggcggtc tggtgaccgc gacccaggac tcctccctgc aggacggcac gctgatctac
     1741 gaggtgaaga tgcgcggcac caacttcccc cccgacggcc ccgtaatgca gaagaagacc
     1801 atgggctggg aggcctccac cgagcgcctg tacccccgcg acggcgtgct gaagggcgag
     1861 atccaccagg ccctgaagct gaaggacggc ggccactacc tggtggagtt caagaccatc
     1921 tacatggcca agaagcccgt gcaactgccc ggctactact acgtggacac caagctggac
     1981 atcacctccc acaacgagga ctacaccagg gtggagcagt acgagcgctc cgagggccgc
     2041 caccacctgt tcctgtacga cggcagcacc ggcagcggca gctccggcac catggtctcc
     2101 aaaggagaag aagtcattaa agagtttatg cgcttcaaag tccgcatgga aggcagcatg
     2161 aacggccacg agttcgagat cgagggcgag ggcgagggca ggccctacga gggaacacag
     2221 acagctaaac tgaaagtcac aaaaggagga cctctgcctt tcgcttggga tatcctgagc
     2281 ccccagttca tgtacggcag caaggcctac gtgaagcacc ccgccgacat ccctgattat
     2341 aaaaaactgt cctttcctga aggattcaaa tgggaacgcg tcatgaattt cgaggacggc
     2401 ggcctggtga ccgtgaccca ggacagcagc atccaggacg gcaccctgat ctataaagtc
     2461 aaaatgcgcg gaacaaattt ccctcctgat ggacctgtca tgcagaaaaa aaccatgggc
     2521 tgggaagcca gcaccgagag gctgtacccc agggacggcg tgctgaaggg cgaaattcat
     2581 caggctctga aactgaaaga tggaggacat tatctggtcg aattcaaaac aatctacatg
     2641 gccaagaagc ccgtgcagct ccccggctac tactacgtgg acaccaagct ggatattaca
     2701 tcccataatg aagattatac aattgtcgaa cagtatgaac gctccgaagg aaggcaccac
     2761 ctctttctgt acggcatgga cgagctgtac aagtaagctt gcctgttttg gcggatgaga
     2821 gaagattttc agcctgatac agattaaatc agaacgcaga agcggtctga taaaacagaa
     2881 tttgcctggc ggcagtagcg cggtggtccc acctgacccc atgccgaact cagaagtgaa
     2941 acgccgtagc gccgatggta gtgtggggtc tccccatgcg agagtaggga actgccaggc
     3001 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     3061 cggtgaacgc tctcctgagt aggacaaatc cgccgggagc ggatttgaac gttgcgaagc
     3121 aacggcccgg agggtggcgg gcaggacgcc cgccataaac tgccaggcat caaattaagc
     3181 agaaggccat cctgacggat ggcctttttg cgtttctaca aactcttttg tttatttttc
     3241 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     3301 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     3361 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     3421 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     3481 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     3541 tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg ccgcatacac
     3601 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     3661 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     3721 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     3781 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     3841 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     3901 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     3961 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     4021 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     4081 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     4141 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     4201 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     4261 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     4321 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     4381 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     4441 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     4501 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     4561 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     4621 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     4681 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     4741 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     4801 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     4861 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     4921 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     4981 tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     5041 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     5101 gtgagcgagg aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt
     5161 atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc
     5221 cagtatacac tccgctatcg ctacgtgact gggtcatggc tgcgccccga cacccgccaa
     5281 cacccgctga cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg
     5341 tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga
     5401 ggcagcagat caattcgcgc gcgaaggcga agcggcatgc ataatgtgcc tgtcaaatgg
     5461 acgaagcagg gattctgcaa a
//
