LOCUS       sequence-196777-        6822 bp    DNA     circular SYN 14-APR-2025
DEFINITION  expresses cytoplasmic ATP sensor in mammalian cells with red 
            reference protein.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6822)
  AUTHORS   Lobas MA, Tao R, Nagai J, Kronschlager MT, Borden PM, Marvin JS, 
            Looger LL, Khakh BS
  TITLE     A genetically encoded single-wavelength sensor for imaging cytosolic
            and cell surface ATP.
  JOURNAL   Nat Commun. 2019 Feb 12;10(1):711. doi: 10.1038/s41467-019-08441-5.
  PUBMED    30755613
REFERENCE   2  (bases 1 to 6822)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2019-02-12"; volume: "10(1):711. doi"; pages: " 
            10.1038/s41467-019-08441-5"
COMMENT     Sequence Label: cyto-Ruby3-iATPSnFR1.0
FEATURES             Location/Qualifiers
     source          1..6822
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     1945..1966
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(2069..2090)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2564..2581)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2570..2794
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2926..3514)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3006..3025)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3843..3890
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3915..3934
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4122..4916)
                     /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
                     GLAPAELFARLKASKPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4233..4252)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4843..4862
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5015..5034)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5113..5304
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5261..5281
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(5364..6155,6156..6224))
                     /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     5987..6006
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6225..6329)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6356..6811
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6443..6462)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6653..6674
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag aactctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgaac tcggatccac tagtaacggc cgccagtgtg ctggaattca tggtgtctaa
      721 gggcgaagag ctgatcaagg aaaatatgcg tatgaaggtg gtcatggaag gttcggtcaa
      781 cggccaccaa ttcaaatgca caggtgaagg agaaggcagg ccgtacgagg gagtgcaaac
      841 catgaggatc aaagtcatcg agggaggacc cctgccattt gcctttgaca ttcttgccac
      901 gtcgttcatg tatggcagcc gtacttttat caagtacccg gcagacattc ctgatttctt
      961 taaacagtcc tttcctgagg gttttacttg ggaaagagtt acgagatacg aagatggtgg
     1021 agtcgtcacc gtcacgcagg acaccagcct tgaggatggc gaactcgttt acaacgtcaa
     1081 agtcagaggg gtaaactttc cctccaatgg tcccgtgatg cagaagaaga ccaagggttg
     1141 ggagcctaat acagagatga tgtatccagc agatggtggt ctgaggggat acactgatat
     1201 tgcactgaaa gttgatggtg gtggccatct gcattgcaat ttcgtaacaa cttacaggtc
     1261 aaaaaagacc gtcgggaaca tcaagatgcc cggtgtccat gccgttgatc accgcctgga
     1321 aaggatagag gaaagtgaca atgaaacgta cgtagtacaa cgcgaagtcg cagttgccaa
     1381 gtactccaat cttggtggtg ctagcatgaa gactattcac gtgagtgtcg taactcccga
     1441 cgggcctgta tatgaggatg acgttgaaat ggtgagcgtc aaagcaaaaa gtggcgagct
     1501 cggtattctc ccaggccaca ttcccttgaa agctccccta gagatcagtg ccgcacgcct
     1561 gaagaaaggg ggcaaaacac agtatatcgc tgtgtcaggc ggcaacctcg aagtgcggcc
     1621 tgacaaggtg accatctatg cccaggcagc cgagagggct gaggatatcg atgtcctgcg
     1681 cgccaaggcc gccaaggaga gagccgagcg ccgactccaa tcacaggtcc tgagccacaa
     1741 cgtctatatc accgccgaca agcagaagaa cggcatcaag gcgaacttca agatccgcca
     1801 caacgtggag gacggcagca tgcagctcgc cgaccactac cagcagaaca cccccatcgg
     1861 cgacggcccc gtgctgctgc ccgacaacca ctacctgagc acccagtccg tgctgagcaa
     1921 agaccctaac gagaagcgcg atcacatggt cctgctggag ttcgtgaccg ccgccgggat
     1981 cactctcggc atggacgagc tgtacaaggg cggtaccgga gggagcatga gcaagggcga
     2041 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     2101 caagttcagc gtgcgcggcg agggcgaggg cgatgccacc aacggcaagc tgaccctgaa
     2161 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
     2221 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     2281 gtccgccatg cccgaaggct acgtccagga gcgcaccatc agcttcaagg acgacggcac
     2341 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     2401 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaactt
     2461 tgggttgcac gacatcgatt ttaagcgcgc cgagctcgcc cttaagcgcg caatgaatag
     2521 gctctcagtt gccgaaatga agtaggcggc cgctcgagat cagcctcgac tgtgccttct
     2581 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2641 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2701 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2761 agcaggcgtg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagtggc
     2821 ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg
     2881 ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg
     2941 cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg
     3001 actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac
     3061 cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca
     3121 tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt
     3181 gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc
     3241 caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag
     3301 agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac
     3361 tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt
     3421 tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa
     3481 gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg
     3541 gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa
     3601 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     3661 atatgagtaa cctgaggcta tggcagggcc tgccgccccg acgttggctg cgagccctgg
     3721 gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa gaaacgcggg cgtattggcc
     3781 ccaatggggt ctcggtgggg tatcgacaga gtgccagccc tgggaccgaa ccccgcgttt
     3841 atgaacaaac gacccaacac cgtgcgtttt attctgtctt tttattgccg tcatagcgcg
     3901 ggttccttcc ggtattgtct ccttccgtgt ttcagttagc ctccccctag ggtgggcgaa
     3961 gaactccagc atgagatccc cgcgctggag gatcatccag ccggcgtccc ggaaaacgat
     4021 tccgaagccc aacctttcat agaaggcggc ggtggaatcg aaatctcgtg atggcaggtt
     4081 gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg ctcagaagaa ctcgtcaaga
     4141 aggcgataga aggcgatgcg ctgcgaatcg ggagcggcga taccgtaaag cacgaggaag
     4201 cggtcagccc attcgccgcc aagctcttca gcaatatcac gggtagccaa cgctatgtcc
     4261 tgatagcggt ccgccacacc cagccggcca cagtcgatga atccagaaaa gcggccattt
     4321 tccaccatga tattcggcaa gcaggcatcg ccatgggtca cgacgagatc ctcgccgtcg
     4381 ggcttgctcg ccttgagcct ggcgaacagt tcggctggcg cgagcccctg atgctcttcg
     4441 tccagatcat cctgatcgac aagaccggct tccatccgag tacgtgctcg ctcgatgcga
     4501 tgtttcgctt ggtggtcgaa tgggcaggta gccggatcaa gcgtatgcag ccgccgcatt
     4561 gcatcagcca tgatggatac tttctcggca ggagcaaggt gagatgacag gagatcctgc
     4621 cccggcactt cgcccaatag cagccagtcc cttcccgctt cagtgacaac gtcgagcaca
     4681 gctgcgcaag gaacgcccgt cgtggccagc cacgatagcc gcgctgcctc gtcttgcagt
     4741 tcattcaggg caccggacag gtcggtcttg acaaaaagaa ccgggcgccc ctgcgctgac
     4801 agccggaaca cggcggcatc agagcagccg attgtctgtt gtgcccagtc atagccgaat
     4861 agcctctcca cccaagcggc cggagaacct gcgtgcaatc catcttgttc aatcatgcga
     4921 aacgatcctc atcctgtctc ttgatcgatc tttgcaaaag cctaggcctc caaaaaagcc
     4981 tcctcactac ttctggaata gctcagaggc cgggcctcgg cctctgcata aataaaaaaa
     5041 attagtcagc catggggcgg agaatgggcg gaactgggcg gagttagggg cgggatgggc
     5101 ggagttaggg gcgggactat ggttgctgac taattgagat gcttgctttg catacttctg
     5161 cctgctgggg agcctgggga ctttccacac ctggttgctg actaattgag atgcttgctt
     5221 tgcatacttc tgcctgctgg ggagcctggg gactttccac accctaactg acacacattc
     5281 cacagctggt tctttccgcc tcaggactct tcctttttca ataaatcaat ctaaagtata
     5341 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     5401 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     5461 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     5521 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     5581 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     5641 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     5701 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     5761 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     5821 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     5881 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     5941 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6001 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6061 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6121 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6181 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     6241 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     6301 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgcg
     6361 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     6421 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     6481 gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct
     6541 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     6601 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     6661 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     6721 attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     6781 aattttaaca aaatattaac gcttacaatt tacgcgcgcg tt
//
