LOCUS       sequence-373043-        7266 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian 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 7266)
  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 7266)
  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/pcDNA3
FEATURES             Location/Qualifiers
     source          1..7266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     43..60
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    177..198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(213..218,219..236,237..243)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    251..267
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     256..278
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     275..291
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     275..291
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    340..461
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(351..370)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     405..424
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(635..1429)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(746..765)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     1356..1375
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1496..1825)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1510..1645
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(1564..1583)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1810..1830
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(1839..2267)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1949..1970)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2161..2180
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    2313..2537
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     2526..2543
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     promoter        2563..2581
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2563..2580
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             complement(2645..3379)
                     /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"
     CDS             complement(join(3959..4471,4472..4474,4475..4477))
                     /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"
     primer_bind     complement(4518..4537)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4519..4537)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(4582..4785)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4611..4631)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4786..5165
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     5337..5356
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        5431..5535
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5536..5604,5605..6396)
                     /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(5754..5773)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6567..7155
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7056..7075
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
       61 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
      121 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
      181 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
      241 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
      301 gccaagctct agctagaggt cgacggtata cagacatgat aagatacatt gatgagtttg
      361 gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta
      421 ttgctttatt tgtaaccatt ataagctgca ataaacaagt tggggtgggc gaagaactcc
      481 agcatgagat ccccgcgctg gaggatcatc cagccggcgt cccggaaaac gattccgaag
      541 cccaaccttt catagaaggc ggcggtggaa tcgaaatctc gtgatggcag gttgggcgtc
      601 gcttggtcgg tcatttcgaa ccccagagtc ccgctcagaa gaactcgtca agaaggcgat
      661 agaaggcgat gcgctgcgaa tcgggagcgg cgataccgta aagcacgagg aagcggtcag
      721 cccattcgcc gccaagctct tcagcaatat cacgggtagc caacgctatg tcctgatagc
      781 ggtccgccac acccagccgg ccacagtcga tgaatccaga aaagcggcca ttttccacca
      841 tgatattcgg caagcaggca tcgccatggg tcacgacgag atcctcgccg tcgggcatgc
      901 gcgccttgag cctggcgaac agttcggctg gcgcgagccc ctgatgctct tcgtccagat
      961 catcctgatc gacaagaccg gcttccatcc gagtacgtgc tcgctcgatg cgatgtttcg
     1021 cttggtggtc gaatgggcag gtagccggat caagcgtatg cagccgccgc attgcatcag
     1081 ccatgatgga tactttctcg gcaggagcaa ggtgagatga caggagatcc tgccccggca
     1141 cttcgcccaa tagcagccag tcccttcccg cttcagtgac aacgtcgagc acagctgcgc
     1201 aaggaacgcc cgtcgtggcc agccacgata gccgcgctgc ctcgtcctgc agttcattca
     1261 gggcaccgga caggtcggtc ttgacaaaaa gaaccgggcg cccctgcgct gacagccgga
     1321 acacggcggc atcagagcag ccgattgtct gttgtgccca gtcatagccg aatagcctct
     1381 ccacccaagc ggccggagaa cctgcgtgca atccatcttg ttcaatcatg cgaaacgatc
     1441 ctcatcctgt ctcttgatca gatccgaaaa tggatataca agctcccggg agctttttgc
     1501 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg
     1561 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
     1621 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
     1681 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
     1741 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
     1801 actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta
     1861 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     1921 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     1981 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     2041 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     2101 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     2161 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     2221 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtggg gataccccct
     2281 agagccccag ctggttcttt ccgcctcaga agccatagag cccaccgcat ccccagcatg
     2341 cctgctattg tcttcccaat cctccccctt gctgtcctgc cccaccccac cccccagaat
     2401 agaatgacac ctactcagac aatgcgatgc aatttcctca ttttattagg aaaggacagt
     2461 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
     2521 gcaactagaa ggcacagtcg aggctgatca gcgagctcta gcatttaggt gacactatag
     2581 aatagggccc tctagatgca tgctcgagcg gccgccagtg tgatggatat ctgcagaatt
     2641 cttactcgat gttgtggcgg atcttgaagt tggccttgat gccgttcttc tgcttgtcgg
     2701 cggtgatata gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc
     2761 cgtcctcctt gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga
     2821 acttcacctc ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct
     2881 ggacgtagcc ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt
     2941 agcgggcgaa gcactgcagg ccgtagccca gggtggtcac gagggtgggc cagggcacgg
     3001 gcagcttgcc ggtggtgcag atcagcttca gggtcagctt gccgtaggtg gcatcgccct
     3061 cgccctcgcc ggacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga
     3121 tgggcaccac cccggtgaac agctcctcgc ccttgctcac catgccaccg ctgccaccct
     3181 tgtacagctc gtccatgccg agagtgatcc cggcggcggt cacgaactcc agcaggacca
     3241 tgtgatcgcg cttctcgttg gggtctttgc tcagggcgga ctggtagctc aggtagtggt
     3301 tgtcgggcag cagcacgggg ccgtcgccga tgggggtgtt ctgctggtag tggtcggcga
     3361 gctgcacgcc gccgtccatg aattcgtttg ccttcccagc cacgtccagt ctgttcaggg
     3421 ccctctgcag tgccagctcg gcccggcgaa tatcggtgtc atcggattga gagttcagcc
     3481 gctcctgggc cctctgtctt gcggcttctg ctctctcttt gtcgattcct tcggctgttt
     3541 cagcggcttg agcattaatg gtaacgtggt caggccgcac ttcaacagtg ccgcctgaga
     3601 ctgcgaccat ctcggtttgg ccgtccttct tcagccgcac agctccgatc ttcagtgggg
     3661 cctttgtggg aatgtgaccg gggaggatgc cgagatcacc actctcggcc cgcacggaca
     3721 ccatctcgat atcagcgtcg tagactggcc catcaggggt tgttatattc actttcacag
     3781 ttttcatatc gatggcggcg gtcacgaact ccagcaggac catgtgatcg cgcttctcgt
     3841 tggggtcttt gctcagggcg gactgggtgc tcaggtagtg gttgtcgggc agcagcacgg
     3901 ggccgtcgcc gatgggggtg ttctgctggt agtggtcggc gagctgcacg ccgccgtcct
     3961 cgatgttgtg gcggatcttg aagtgggcct tgatgccgtt cttctgcttg tcggcggtga
     4021 tatagacgtt gtggctgatg tagttgtact ccagcttgtg ccccaggatg ttgccgtcct
     4081 ccttgaagtc gatgcccttc agctcgatgc ggttcaccag ggtgtcgccc tcgaacttca
     4141 cctcggcgcg ggtcttgtag ttgccgtcgt ccttgaagaa gatggtacgc tcctggacgt
     4201 agccttcggg catggcggac ttgaagaagt cgtgctgctt catgtggtcg gggtagcggg
     4261 cgaagcactg cacgccccag gtcagggtgg tcacgagggt gggccagggc acgggcagct
     4321 tgccggtggt gcagatgaac ttcagggtca gcttgccgta ggtggcatcg ccctcgccct
     4381 cgccggacac gctgaacctg tggccgttta cgtcgccgtc cagctcgacc aggatgggca
     4441 ccaccccggt gaacagctcc tcgcccttgc tcaccatact cgagctcgga tccgagctcg
     4501 gtaccaagct tgggtctccc tatagtgagt cgtattaatt tcgataagcc agtaagcagt
     4561 gggttctcta gttagccaga gagctctgct tatatagacc tcccaccgta cacgcctacc
     4621 gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc gttgattttg
     4681 gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc
     4741 aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg taatagcgat
     4801 gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt actgggcata
     4861 atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg gcatatgata
     4921 cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt gacgtcaatg
     4981 gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca atgggcgggg
     5041 gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc ggaactccat
     5101 atatgggcta tgaactaatg accccgtaat tgattactat taataactag tcaataatca
     5161 atgtcaacgc gtatatctgg cccgtacatc gcgaagcagc gcaaaacgcc taaccctaag
     5221 cagattcttc atgcaattgt cggtcaagcc ttgccttgtt gtagcttaaa ttttgctcgc
     5281 gcactactca gcgacctcca acacacaagc agggagcaga tactggctta actatgcggc
     5341 atcagagcag attgtactga gagtgcacca taggggatcg ggagatctcc cgatccgtcg
     5401 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     5461 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     5521 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     5581 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     5641 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     5701 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     5761 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     5821 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     5881 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     5941 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     6001 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     6061 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     6121 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     6181 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     6241 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     6301 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     6361 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     6421 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     6481 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     6541 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     6601 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     6661 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     6721 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     6781 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     6841 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     6901 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     6961 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     7021 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     7081 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     7141 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     7201 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     7261 gccttt
//
