LOCUS       sequence-429941-        4696 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  bacterial expression of ATeam1.03-nD/nA, a FRET-based ATP indicator.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4696)
  AUTHORS   Kotera I, Iwasaki T, Imamura H, Noji H, Nagai T
  TITLE     Reversible dimerization of Aequorea victoria fluorescent proteins 
            increases the dynamic range of FRET-based indicators.
  JOURNAL   ACS Chem Biol. 2010 Feb 19;5(2):215-22. doi: 10.1021/cb900263z.
  PUBMED    20047338
REFERENCE   2  (bases 1 to 4696)
  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"; doi: "10"; journalName: 
            "ACS Chem Biol"; date: "2010-02-19- 19"; volume: "5"; issue: "2"; 
            pages: "215-22"
COMMENT     Sequence Label: Ateam1.03-nD/nA/pRSETB
FEATURES             Location/Qualifiers
     source          1..4696
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /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(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             join(189..191,192..194,195..707)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHRFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     AHFKIRHNIE"
     CDS             1287..2021
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRD
                     HMVLLEFVTAAGITLGMDELYKGGSGGMVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKIRHNIE"
     primer_bind     complement(2078..2096)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2092..2139
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2236..2691
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2323..2342)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2533..2554
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2717..2821
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2822..2890,2891..3682)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3040..3059)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3853..4441
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4342..4361
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4595..4612
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatccga
      181 gctcgagtat ggtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg
      241 agctggacgg cgacgtaaac ggccacaggt tcagcgtgtc cggcgagggc gagggcgatg
      301 ccacctacgg caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct
      361 ggcccaccct cgtgaccacc ctgacctggg gcgtgcagtg cttcgcccgc taccccgacc
      421 acatgaagca gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgta
      481 ccatcttctt caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg
      541 acaccctggt gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc
      601 tggggcacaa gctggagtac aactacatca gccacaacgt ctatatcacc gccgacaagc
      661 agaagaacgg catcaaggcc cacttcaaga tccgccacaa catcgaggac ggcggcgtgc
      721 agctcgccga ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg
      781 acaaccacta cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc
      841 acatggtcct gctggagttc gtgaccgccg ccatcgatat gaaaactgtg aaagtgaata
      901 taacaacccc tgatgggcca gtctacgacg ctgatatcga gatggtgtcc gtgcgggccg
      961 agagtggtga tctcggcatc ctccccggtc acattcccac aaaggcccca ctgaagatcg
     1021 gagctgtgcg gctgaagaag gacggccaaa ccgagatggt cgcagtctca ggcggcactg
     1081 ttgaagtgcg gcctgaccac gttaccatta atgctcaagc cgctgaaaca gccgaaggaa
     1141 tcgacaaaga gagagcagaa gccgcaagac agagggccca ggagcggctg aactctcaat
     1201 ccgatgacac cgatattcgc cgggccgagc tggcactgca gagggccctg aacagactgg
     1261 acgtggctgg gaaggcaaac gaattcatgg acggcggcgt gcagctcgcc gaccactacc
     1321 agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagct
     1381 accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt
     1441 tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagggt ggcagcggtg
     1501 gcatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
     1561 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
     1621 acggcaagct gaccctgaag ctgatctgca ccaccggcaa gctgcccgtg ccctggccca
     1681 ccctcgtgac caccctgggc tacggcctgc agtgcttcgc ccgctacccc gaccacatga
     1741 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
     1801 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1861 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1921 acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac aagcagaaga
     1981 acggcatcaa ggccaacttc aagatccgcc acaacatcga gtaagcttga tccggctgct
     2041 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     2101 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     2161 ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     2221 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     2281 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     2341 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     2401 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     2461 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     2521 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     2581 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     2641 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     2701 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     2761 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     2821 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     2881 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     2941 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     3001 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     3061 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     3121 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     3181 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     3241 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     3301 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     3361 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     3421 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     3481 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     3541 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     3601 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     3661 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     3721 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     3781 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     3841 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     3901 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     3961 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     4021 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4081 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4141 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4201 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4261 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4321 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4381 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4441 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     4501 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     4561 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     4621 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcaggat
     4681 ctcgatcccg cgaaat
//
