LOCUS       sequence-254288-        7756 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Express Positron in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7756)
  AUTHORS   Abdelfattah AS, Valenti R, Zheng J, Wong A, Podgorski K, Koyama M, 
            Kim DS, Schreiter ER
  TITLE     A general approach to engineer positive-going eFRET voltage 
            indicators.
  JOURNAL   Nat Commun. 2020 Jul 10;11(1):3444. doi: 10.1038/s41467-020-17322-1.
  PUBMED    32651384
REFERENCE   2  (bases 1 to 7756)
  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: "Nat 
            Commun."; date: "2020-07-10"; volume: "11(1):3444. doi"; pages: " 
            10.1038/s41467-020-17322-1"
COMMENT     Sequence Label: pCAG-Positron
FEATURES             Location/Qualifiers
     source          1..7756
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      8..17
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1787..1804)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1793..2017
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2063..2491
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2150..2169)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2360..2381
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2500..2520)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2505..2834
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2685..2820
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2747..2766
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2845..3444
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2845..2864)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3341..3361
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3604..3725
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3641..3660)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3695..3714
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3774..3790)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3774..3790)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        complement(join(3822..3828,3829..3846,3847..3852))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3867..3888
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4005..4022)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4176..4764)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4256..4275)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4935..5726,5727..5795))
                     /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     5558..5577
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5796..5900)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        5932..6311
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          6588..7604
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7528..7550
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     7612..7631
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
ORIGIN
        1 gctagccgcc accatggctg acgtggaaac cgagaccggc atgattgcac agtggattgt
       61 ctttgctatt atggctgctg ctgctattgc ttttggagtg gctgtgcact ttcggccttc
      121 agagctgaag agcgcatact atatcaacat tgccatctgc actatcgccg ctaccgctta
      181 ctatgcaatg gccgtgaact accaggacct gacaatgaat ggtgaaaggc aggtggtcta
      241 cgcaagatat attgactggg tgctgaccac accactgctc ctgctcaacc tcatcgtcat
      301 gaccaagatg ggcggagtga tgatttcttg ggtcatcggc gcagacattt tcatgatcgt
      361 gtttggtatt ctgggcgcct tcgaggatga acacaagttc aaatgggtgt actttatcgc
      421 tggatgtgtg atgcaggcag tcctgacata cgggatgtat aacgccactt ggaaagacga
      481 tctgaagaaa agccccgagt accatagctc ctatgtcagt ctgctcgtct tcctgtcaat
      541 cctctgggtg ttttatcctg tcgtgtgggc tttcgggtct ggtagtggcg tgctgtccgt
      601 cgacaatgtg gccattctca tgggaatcct ggatgtgctc gctaagccac tgtttggaat
      661 ggggtgcctc attgcccatg agactatctt caagatcggt actggctttc cattcgaccc
      721 ccattatgtg gaagtcctgg gcgagcgcat gcactacgtc gatgttggtc cgcgcgatgg
      781 cacccctgtg ctgttcctgc acggtaaccc gacctcctcc tacgtgtggc gcaacatcat
      841 cccgcatgtt gcaccgaccc atcgctgcat tgctccagac ctgatcggta tgggcaaatc
      901 cgacaaacca gacctgggtt atttcttcga cgaccacgtc cgcttcatgg atgccttcat
      961 cgaagccctg ggtctggaag aggtcgtcct ggtcattcac gactggggct ccgctctggg
     1021 tttccactgg gccaagcgca atccagagcg cgtcaaaggt attgcattta tggagttcat
     1081 ccgccctatc ccgacctggg acgaatggcc agaatttgcc cgcgagacct tccaggcctt
     1141 ccgcaccacc gacgtcggcc gcaagctgat catcgatcag aacgttttta tcgagggtac
     1201 gctgccgatg ggtgtcgtcc gcccgctgac tgaagtcgag atggaccatt accgcgagcc
     1261 gttcctgaat cctgttgacc gcgagccact gtggcgcttc ccaaacgagc tgccaatcgc
     1321 cggtgagcca gcgaacatcg tcgcgctggt cgaagaatac atggactggc tgcaccagtc
     1381 ccctgtcccg aagctgctgt tctggggcac cccaggcgtt ctgatcccac cggccgaagc
     1441 cgctcgcctg gccaaaagcc tgcctaactg caaggctgtg gacatcggcc cgggtctgaa
     1501 tctgctgcaa gaagacaacc cggacctgat cggcagcgag atcgcgcgct ggctgtcgac
     1561 gctcgagatt tccggcgagc caaccactaa gagcaggatc accagcgagg gcgagtacat
     1621 ccccctggac cagatcgaca tcaacgtgtt ctgctacgag aacgaggtgt aatgaaagct
     1681 tggtaccgag ctcggatcca ctagtccagt gtggtggaat tctgcagata tccagcacag
     1741 tggcggccgc tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     1801 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1861 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     1921 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     1981 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     2041 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     2101 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     2161 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     2221 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     2281 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     2341 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     2401 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     2461 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     2521 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     2581 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     2641 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     2701 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     2761 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     2821 taggcttttg caaaaagctc ccgcatgacc gagtacaagc ccacggtgcg cctcgccacc
     2881 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     2941 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     3001 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     3061 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     3121 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     3181 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     3241 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     3301 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     3361 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     3421 atgacccgca agcccggtgc ctgattcgaa atgaccgacc aagcgacgcc caacctgcca
     3481 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3541 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3601 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3661 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3721 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3781 ctgtttcctg tgtgaaatta ttatccgctc acaattccac acaacatacg agccggaagc
     3841 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3901 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3961 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4021 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4081 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4141 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4201 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4261 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4321 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4381 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4441 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4501 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4561 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4621 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4681 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4741 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4801 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4861 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4921 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4981 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5041 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5101 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5161 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5221 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5281 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5341 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5401 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5461 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5521 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5581 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5641 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5701 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5761 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5821 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5881 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacattgat
     5941 tattgactag ttattaatag taatcaatta cggggtcatt agttcatagc ccatatatgg
     6001 agttccgcgt tacataactt acggtaaatg gcccgcctgg ctgaccgccc aacgaccccc
     6061 gcccattgac gtcaataatg acgtatgttc ccatagtaac gccaataggg actttccatt
     6121 gacgtcaatg ggtggagtat ttacggtaaa ctgcccactt ggcagtacat caagtgtatc
     6181 atatgccaag tacgccccct attgacgtca atgacggtaa atggcccgcc tggcattatg
     6241 cccagtacat gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg
     6301 ctattaccat ggtcgaggtg agccccacgt tctgcttcac tctccccatc tcccccccct
     6361 ccccaccccc aattttgtat ttatttattt tttaattatt ttgtgcagcg atgggggcgg
     6421 gggggggggg nggsgcgccr ggsggggsgg ggsggggsgr ggggsggggs ggggsgaggc
     6481 ggagaggtgc ggcggcagcc aatcagagcg gcgcgctccg aaagtttcct tttatggcga
     6541 ggcggcggcg gcggcggccc tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc
     6601 gctgccttcg ccccgtgccc cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac
     6661 tgaccgcgtt actcccacag gtgagcgggc gggacggccc ttctcctccg ggctgtaatt
     6721 agcgcttggt ttaatgacgg cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc
     6781 tccgggaggg ccctttgtgc ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg
     6841 tggggagcgc cgcgtgcggc tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg
     6901 cggggctttg tgcgctccgc agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc
     6961 ggtgcggggg gggctgcgag gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt
     7021 gagcaggggg tgtgggcgcg tcggtcgggc tgcaaccccc cctgcacccc cctccccgag
     7081 ttgctgagca cggcccggct tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg
     7141 ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg
     7201 ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg
     7261 cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag ggacttcctt
     7321 tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc
     7381 gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc cttcgtgcgt
     7441 cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg ggggacggct
     7501 gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc ggcggctcta
     7561 gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg ggcaacgtgc
     7621 tggttattgt gctgtctcat cattttggca aagaattcct cgaggaattc actcctcagg
     7681 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
     7741 actgagatct gcatga
//
