LOCUS       sequence-478621-        7792 bp    DNA     circular SYN 15-JUL-2025
DEFINITION  Mammalian expression of positive-going voltage sensor.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7792)
  AUTHORS   Eric R. Schreiter, Ahmed Abdelfattah, Jeremy Hasseman, Daniel Reep, 
            Getahun Tsegaye, Arthur Tsang, Ilya Kolb, Allan Wong, Amy Chuong, 
            Jihong Zheng, Ben Arthur, Wyatt Korff, GENIE Project Team
  TITLE     Optimization of Ace2N based voltage indicators
  JOURNAL   Janelia Figshare 2023
REFERENCE   2  (bases 1 to 7792)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Janelia 
            Figshare 2023"
COMMENT     Sequence Label: pGP-pcDNA3.1 Puro-CAG-Positron2
FEATURES             Location/Qualifiers
     source          1..7792
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     57..76
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     regulatory      209..218
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(1988..2005)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1994..2218
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2264..2692
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2351..2370)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2561..2582
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2701..2721)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2706..3035
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2886..3021
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2948..2967
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3046..3645
                     /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(3046..3065)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3542..3562
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3805..3926
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3842..3861)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3896..3915
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3975..3991)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3975..3991)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3988..4010)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3999..4015
                     /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(4023..4029,4030..4047,4048..4053))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4068..4089
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4206..4223)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4377..4965)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4457..4476)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5136..5927,5928..5996))
                     /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     5759..5778
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5997..6101)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        6133..6512
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     intron          join(6825..7792,1..49)
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     7765..7787
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
ORIGIN
        1 ctctagagcc tctgctaacc atgttcatgc cttcttcttt ttcctacagc tcctgggcaa
       61 cgtgctggtt attgtgctgt ctcatcattt tggcaaagaa ttcctcgagg aattcactcc
      121 tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc tggctcacaa
      181 ataccactga gatctgcatg agctagccgc caccatggct gacgtggaaa ccgagaccgg
      241 tatgattgca cagtggattg tctttgctat tatggctgct gctgctattg cttttggagt
      301 ggctgtgcac tttcggcctt cagagctgaa gagcgcatac tatatcaaca ttgccatctg
      361 cactatcgcc gctaccgctt actatgcaat ggccgtgaac taccaggacc tgacaatgaa
      421 tggtgaaagg caggtggtct acgcaaaata tattgactgg gtgctgacca caccactgct
      481 cctgctcaac ctcatcgtca tgaccaagat gggcggagtg atgatttctt gggtcatcgg
      541 cgcagacatt ttcatgatcg tgtttggtat tctgggcgcc ttcgaggatg aacacaagtt
      601 caaatgggtg tactttatcg ctggatgtgt gatgcaggca gtcctgacat acgggatgta
      661 taacgccact tggaaagacg atctgaagaa aagccccgag taccatagct cctatgtcag
      721 tctgctcgtc ttcctgtcaa tcctcttcgt gttttatcct gtcgtgtggg ctttcgggtc
      781 tggtagtggc gtgctgtccg tcgacaatga ggccattctc atgggaatcc tggatgtgct
      841 cgctaagcca ctgtttggaa tggggtgcct cattgcccat gagactatct tcaagatcgg
      901 taccggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca tgcactacgt
      961 cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc cgacctcctc
     1021 ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca ttgctccaga
     1081 cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg acgaccacgt
     1141 ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc tggtcattca
     1201 cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc gcgtcaaagg
     1261 tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc cagaatttgc
     1321 ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga tcatcgatca
     1381 gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga ctgaagtcga
     1441 gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac tgtggcgctt
     1501 cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg tcgaagaata
     1561 catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca ccccaggcgt
     1621 tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact gcaaggctgt
     1681 ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga tcggcagcga
     1741 gatcgcgcgc tggctgtcga cgctcgagat ttccggcgag ccaaccacta agagcaggat
     1801 caccagcgag ggcgagtaca tccccctgga ccagatcgac atcaacgtgt tctgctacga
     1861 gaacgaggtg taatgaaagc ttcgtaccga gctcggatcc actagtccag tgtggtggaa
     1921 ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt ttaaacccgc
     1981 tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg
     2041 ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt
     2101 gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc
     2161 aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct
     2221 tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc
     2281 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     2341 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     2401 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     2461 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     2521 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     2581 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     2641 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     2701 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2761 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     2821 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     2881 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     2941 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3001 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgcatgac cgagtacaag
     3061 cccacggtgc gcctcgccac ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc
     3121 gcgttcgccg actaccccgc cacgcgccac accgtcgatc cggaccgcca catcgagcgg
     3181 gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg
     3241 gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg
     3301 gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga gcggttcccg gctggccgcg
     3361 cagcaacaga tggaaggcct cctggcgccg caccggccca aggagcccgc gtggttcctg
     3421 gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc
     3481 cccggagtgg aggcggccga gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc
     3541 cgcaacctcc ccttctacga gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc
     3601 gaaggaccgc gcacctggtg catgacccgc aagcccggtg cctgattcga aatgaccgac
     3661 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3721 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3781 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3841 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3901 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3961 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4021 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4081 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4141 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4201 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4261 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4321 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4381 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4441 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     4501 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     4561 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4621 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4681 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4741 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4801 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4861 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4921 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4981 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5041 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5101 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5161 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5221 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5281 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5341 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5401 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5461 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5521 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5581 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5641 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5701 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5761 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5821 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5881 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5941 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6001 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6061 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6121 ccacctgacg tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat
     6181 tagttcatag cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg
     6241 gctgaccgcc caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa
     6301 cgccaatagg gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact
     6361 tggcagtaca tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta
     6421 aatggcccgc ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt
     6481 acatctacgt attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttca
     6541 ctctccccat ctcccccccc tccccacccc caattttgta tttatttatt ttttaattat
     6601 tttgtgcagc gatgggggcg gggggggggg gggggcgcgc cagggggggg gggggggggg
     6661 gggggggggg gggggggggg gggggggggg ggggggaggg gcggggcggg gcgaggcgga
     6721 gaggtgcggc ggcagccaat cagagcggcg cgctccgaaa gtttcctttt atggcgaggc
     6781 ggcggcggcg gcggccctat aaaaagcgaa gcgcgcggcg ggcgggagtc gctgcgcgct
     6841 gccttcgccc cgtgccccgc tccgccgccg cctcgcgccg cccgccccgg ctctgactga
     6901 ccgcgttact cccacaggtg agcgggcggg acggcccttc tcctccgggc tgtaattagc
     6961 gcttggttta atgacggctt gtttcttttc tgtggctgcg tgaaagcctt gaggggctcc
     7021 gggagggccc tttgtgcggg gggagcggct cggggggtgc gtgcgtgtgt gtgtgcgtgg
     7081 ggagcgccgc gtgcggctcc gcgctgcccg gcggctgtga gcgctgcggg cgcggcgcgg
     7141 ggctttgtgc gctccgcagt gtgcgcgagg ggagcgcggc cgggggcggt gccccgcggt
     7201 gcgggggggg ctgcgagggg aacaaaggct gcgtgcgggg tgtgtgcgtg ggggggtgag
     7261 cagggggtgt gggcgcgtcg gtcgggctgc aaccccccct gcacccccct ccccgagttg
     7321 ctgagcacgg cccggcttcg ggtgcggggc tccgtacggg gcgtggcgcg gggctcgccg
     7381 tgccgggcgg ggggtggcgg caggtggggg tgccgggcgg ggcggggccg cctcgggccg
     7441 gggagggctc gggggagggg cgcggcggcc cccggagcgc cggcggctgt cgaggcgcgg
     7501 cgagccgcag ccattgcctt ttatggtaat cgtgcgagag ggcgcaggga cttcctttgt
     7561 cccaaatctg tgcggagccg aaatctggga ggcgccgccg caccccctct agcgggcgcg
     7621 gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt cgtgcgtcgc
     7681 cgcgccgccg tccccttctc cctctccagc ctcggggctg tccgcggggg gacggctgcc
     7741 ttcggggggg acggggcagg gcggggttcg gcttctggcg tgtgaccggc gg
//
