LOCUS       sequence-381768-        7018 bp    DNA     circular SYN 14-APR-2025
DEFINITION  CMV overexpression of ChemR-ATP in mammalian cell lines.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7018)
  AUTHORS   Hellweg L, Edenhofer A, Barck L, Huppertz MC, Frei MS, Tarnawski M, 
            Bergner A, Koch B, Johnsson K, Hiblot J
  TITLE     A general method for the development of multicolor biosensors with 
            large dynamic ranges.
  JOURNAL   Nat Chem Biol. 2023 Jun 8. doi: 10.1038/s41589-023-01350-1.
  PUBMED    37291200
REFERENCE   2  (bases 1 to 7018)
  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 Chem 
            Biol. 2023 Jun 8. doi: 10.1038/s41589-023-01350-1."
COMMENT     Sequence Label: pCDNA5/FRT-ChemoR-ATP
FEATURES             Location/Qualifiers
     source          1..7018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             917..1627
                     /codon_start=1
                     /product="monomeric red fluorescent protein derived from 
                     mRuby, with improved photophysical properties (Lam et al., 
                     2012)"
                     /label=mRuby2
                     /translation="MVSKGEELIKENMRMKVVMEGSVNGHQFKCTGEGEGNPYMGTQTM
                     RIKVIEGGPLPFAFDILATSFMYGSRTFIKYPKGIPDFFKQSFPEGFTWERVTRYEDGG
                     VVTVMQDTSLEDGCLVYHVQVRGVNFPSNGPVMQKKTKGWEPNTEMMYPADGGLRGYTH
                     MALKVDGGGHLSCSFVTTYRSKKTVGNIKMPGIHAVDHRLERLEESDNEMFVVQREHAV
                     AKFAGLGGGMDELYK"
     primer_bind     complement(2970..2987)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2976..3200
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3333..3352)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3484..3531
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3539..4561
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYKSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    4691..4812
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4728..4747)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4782..4801
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4861..4877)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4861..4877)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4874..4896)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4885..4901
                     /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)."
     promoter        complement(join(4909..4915,4916..4933,4934..4939))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4954..4975
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5092..5109)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5263..5851)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5343..5362)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6022..6813,6814..6882))
                     /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     6645..6664
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6883..6987)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt accaccatgg tgtctaaggg cgaagagctg atcaaggaaa atatgcgtat
      961 gaaggtggtc atggaaggtt cggtcaacgg ccaccaattc aaatgcacag gtgaaggaga
     1021 aggcaatccg tacatgggaa ctcaaaccat gaggatcaaa gtcatcgagg gaggacccct
     1081 gccatttgcc tttgacattc ttgccacgtc gttcatgtat ggcagccgta cttttatcaa
     1141 gtacccgaaa ggcattcctg atttctttaa acagtccttt cctgagggtt ttacttggga
     1201 aagagttacg agatacgaag atggtggagt cgtcaccgtc atgcaggaca ccagccttga
     1261 ggatggctgt ctcgtttacc acgtccaagt cagaggggta aactttccct ccaatggtcc
     1321 cgtgatgcag aagaagacca agggttggga gcctaataca gagatgatgt atccagcaga
     1381 tggtggtctg aggggataca ctcatatggc actgaaagtt gatggtggtg gccatctgtc
     1441 ttgctctttc gtaacaactt acaggtcaaa aaagaccgtc gggaacatca agatgcccgg
     1501 tatccatgcc gttgatcacc gcctggaaag gttagaggaa agtgacaatg aaatgttcgt
     1561 agtacaacgc gaacacgcag ttgccaagtt cgccgggctt ggtggtggga tggacgagct
     1621 gtacaagggt ggcggcatga aaactgtgaa agtgaatata acaacccctg atgggccagt
     1681 ctacgacgct gatatcgaga tggtgtccgt gcgggccgag agtggtgatc tcggcatcct
     1741 ccccggtcac attcccacaa aggccccact gaagatcgga gctgtgcggc tgaagaagga
     1801 cggccaaacc gagatggtcg cagtctcagg cggcactgtt gaagtgcggc ctgaccacgt
     1861 taccattaat gctcaagccg ctgaaacagc cgaaggaatc gacaaagaga gagcagaagc
     1921 cgcaagacag agggcccagg agcggctgaa ctctcaatcc gatgacaccg atattcgccg
     1981 ggccgagctg gcactgcaga gggccctgaa cagactggac gtggctggga aggcaaacga
     2041 attcggtggc ggtatcggta ctggctttcc attcgacccc cattatgtgg aagtcctggg
     2101 cgagcgcatg cactacgtcg atgttggtcc gcgcgatggc acccctgtgc tgttcctgca
     2161 cggtaacccg acctcctcct acgtgtggcg caacatcatc ccgcatgttg caccgaccca
     2221 tcgctgcatt gctccagacc tgatcggtat gggcaaatcc gacaaaccag acctgggtta
     2281 tttcttcgac gaccacgtcc gcttcatgga tgccttcatc gaagccctgg gtctggaaga
     2341 ggtcgtcctg gtcattcacg actggggctc cgctctgggt ttccactggg ccaagcgcaa
     2401 tccagagcgc gtcaaaggta ttgcatttat ggagttcatc cgccctatcc cgacctggga
     2461 cgaatggcca gaatttgccc gcgagacctt ccaggccttc cgcaccaccg acgtcggccg
     2521 caagctgatc atcgatcaga acgtttttat cgagggtacg ctgccgatgg gtgtcgtccg
     2581 cccgctgact gaagtcgaga tggaccatta ccgcgagccg ttcctgaatc ctgttgaccg
     2641 cgagccactg tggcgcttcc caaacgagct gccaatcgcc ggtgagccag cgaacatcgt
     2701 cgcgctggtc gaagaataca tggactggct gcaccagtcc cctgtcccga agctgctgtt
     2761 ctggggcacc ccaggcgttc tgatcccacc ggccgaagcc gctcgcctgg ccaaaagcct
     2821 gcctaactgc aaggctgtgg acatcggccc gggtctgaat ctgctgcaag aagacaaccc
     2881 ggacctgatc ggcagcgaga tcgcgcgctg gctgtctact ctggagattt ccggttagtc
     2941 tagagggccc gtttaaaccc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3001 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3061 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     3121 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     3181 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3241 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3301 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3361 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     3421 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtac
     3481 ctagaagttc ctattccgaa gttcctattc tctagaaagt ataggaactt ccttggccaa
     3541 aaagcctgaa ctcaccgcga cgtctgtcga gaagtttctg atcgaaaagt tcgacagcgt
     3601 ctccgacctg atgcagctct cggagggcga agaatctcgt gctttcagct tcgatgtagg
     3661 agggcgtgga tatgtcctgc gggtaaatag ctgcgccgat ggtttctaca aagatcgtta
     3721 tgtttatcgg cactttgcat cggccgcgct cccgattccg gaagtgcttg acattgggga
     3781 attcagcgag agcctgacct attgcatctc ccgccgtgca cagggtgtca cgttgcaaga
     3841 cctgcctgaa accgaactgc ccgctgttct gcagccggtc gcggaggcca tggatgcgat
     3901 cgctgcggcc gatcttagcc agacgagcgg gttcggccca ttcggaccgc aaggaatcgg
     3961 tcaatacact acatggcgtg atttcatatg cgcgattgct gatccccatg tgtatcactg
     4021 gcaaactgtg atggacgaca ccgtcagtgc gtccgtcgcg caggctctcg atgagctgat
     4081 gctttgggcc gaggactgcc ccgaagtccg gcacctcgtg cacgcggatt tcggctccaa
     4141 caatgtcctg acggacaatg gccgcataac agcggtcatt gactggagcg aggcgatgtt
     4201 cggggattcc caatacgagg tcgccaacat cttcttctgg aggccgtggt tggcttgtat
     4261 ggagcagcag acgcgctact tcgagcggag gcatccggag cttgcaggat cgccgcggct
     4321 ccgggcgtat atgctccgca ttggtcttga ccaactctat aagagcttgg ttgacggcaa
     4381 tttcgatgat gcagcttggg cgcagggtcg atgcgacgca atcgtccgat ccggagccgg
     4441 gactgtcggg cgtacacaaa tcgcccgcag aagcgcggcc gtctggaccg atggctgtgt
     4501 agaagtactc gccgatagtg gaaaccgacg ccccagcact cgtccgaggg caaaggaata
     4561 gcacgtacta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     4621 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     4681 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     4741 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     4801 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     4861 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     4921 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     4981 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5041 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5101 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5161 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5221 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     5281 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     5341 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     5401 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     5461 ctcacgctgt aggtatctca gttcggtgca ggtcgttcgc tccaagctgg gctgtgtgca
     5521 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     5581 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     5641 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     5701 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     5761 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     5821 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     5881 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     5941 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6001 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     6061 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     6121 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     6181 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     6241 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     6301 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     6361 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     6421 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     6481 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     6541 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     6601 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     6661 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     6721 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     6781 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     6841 aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     6901 ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     6961 gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
