LOCUS       sequence-429687-        7768 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression vector for RTN3E-mScarlet3.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7768)
  AUTHORS   Crapart CC, Scott ZC, Konno T, Sharma A, Parutto P, Bailey DMD, 
            Westrate LM, Avezov E, Koslover EF
  TITLE     Luminal transport through intact endoplasmic reticulum limits the 
            magnitude of localized Ca(2+) signals.
  JOURNAL   Proc Natl Acad Sci U S A. 2024 Mar 26;121(13):e2312172121. doi: 
            10.1073/pnas.2312172121. Epub 2024 Mar 19.
  PUBMED    38502705
REFERENCE   2  (bases 1 to 7768)
  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: "Proc Natl 
            Acad Sci U S A."; date: "2024-03-26"; volume: "121(13):e2312172121. 
            doi"; pages: " 10.1073/pnas.2312172121. Epub 2024 Mar 19"
COMMENT     Sequence Label: pCMV_RTN3E-mScarlet3
FEATURES             Location/Qualifiers
     source          1..7768
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        152..455
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        456..659
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     610..630
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             complement(1171..1182)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     CDS             3836..4522
                     /codon_start=1
                     /product="bright cysteine-free monomeric red fluorescent 
                     protein with fast and complete maturation (Gadella et al., 
                     2023)"
                     /label=mScarlet3
                     /translation="MDSTEAVIKEFMRFKVHMEGSMNGHEFEIEGEGEGRPYEGTQTAK
                     LRVTKGGPLPFSWDILSPQFMYGSRAFTKHPADIPDYWKQSFPEGFKWERVMNFEDGGA
                     VSVAQDTSLEDGTLIYKVKLRGTNFPPDGPVMQKKTMGWEASTERLYPEDVVLKGDIKM
                     ALRLKDGGRYLADFKTTYRAKKPVQMPGAFNIDRKLDITSHNEDYTVVEQYERSVARHS
                     TGGSGGS"
     primer_bind     4234..4253
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4647..4768
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4684..4703)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4738..4757
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4775..5230)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4912..4933)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5124..5143
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5257..5361
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5363..5720
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5363..5554
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5386..5406)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5571..5706
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5755..6549
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             5755..6549
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5809..5828)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6419..6438
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6737..6756)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6781..6828
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7157..7745
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7646..7665
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
       61 ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca attacggggt
      121 cattagttca tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc
      181 ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag
      241 taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc
      301 acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg
      361 gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc
      421 agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca
      481 atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca
      541 atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg
      601 ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctg
      661 gtttagtgaa ccgtcagatc cgctagcgct accggactca gatctcgaga tggcggagcc
      721 gtcggcggcc actcagtccc attccatctc ctcgtcgtcc ttcggagccg agccgtccgc
      781 gcccggcggc ggcgggagcc caggagcctg ccccgccctg gggacgaaga gctgcagctc
      841 ctcctgtgcg gattcctttg tttcttcctc ttcctctcag cctgtatctc tattttcgac
      901 ctcacaagag ggattgagct ctctttgctc tgatgagcca tcttcagaaa ttatgacttc
      961 ttcctttctt tcatcttctg aaatacataa cactggcctt acaatactac atggagaaaa
     1021 aagccatgtg ttagggagcc agcctatttt agccaaagaa ggaaaagacc acttggatct
     1081 tctagatatg aaaaagatgg aaaagcctca ggggaccagc aacaacgtat cagactcttc
     1141 agtttctctt gcagcaggag ttcattgtga ccgtccttct attccagcca gtttcccaga
     1201 gcatcctgct tttctctcaa agaaaattgg tcaagtggaa gagcaaatag ataaagagac
     1261 caagaaccca aatggggtat caagtaggga ggctaaaact gcattggatg ctgatgacag
     1321 attcactttg ctgacagccc agaaaccacc tactgagtac tctaaggtag aaggcattta
     1381 tacatattct ttgtctccat ccaaagtttc aggagatgat gttattgaaa aggattcccc
     1441 tgaatcacca tttgaagtaa ttattgacaa agcagcattt gacaaagaat ttaaagactc
     1501 atataaggag agcacagatg attttggtag ctggtctgtg cacactgata aagaatcatc
     1561 cgaagacatt tcagagacta atgacaagct ttttccactg agaaataaag aggcaggacg
     1621 ttacccaatg tctgcattgc tcagtaggca gttttcacac acaaatgcag cactggaaga
     1681 ggtgtccaga tgcgtgaatg atatgcataa ctttactaac gaaatactga cttgggatct
     1741 ggttccccaa gtgaaacaac agaccgataa atcttctgac tgcatcacaa aaactacagg
     1801 acttgacatg agtgaatata attcagaaat tccagttgta aatcttaaaa ctagcactca
     1861 tcagaaaact cctgtatgtt ctattgatgg gagcactccc atcactaaat caacaggtga
     1921 ttgggcagaa gcatctctcc agcaagaaaa tgctattact ggaaaacctg tacctgactc
     1981 tttgaattcc acaaaagaat tcagtatcaa aggtgtgcaa ggcaatatgc agaaacagga
     2041 tgacacactt gcagaattac ctggatctcc acctgagaaa tgtgactctt tgggttctgg
     2101 agtggccaca gtgaaagtgg ttttacctga tgaccacctg aaagatgaaa tggactggca
     2161 gagctctgca ttgggagaaa tcacagaagc tgatagttct ggtgagtctg atgacacagt
     2221 aatagaggac atcacagcag atacatcatt tgaaaataac aaaattcagg ctgaaaaacc
     2281 tgtttccatt ccaagtgctg ttgtaaaaac aggtgaaaga gaaatcaaag agattcccag
     2341 ttgtgagaga gaagaaaaaa catctaaaaa ctttgaagaa ttggtcagtg actctgagct
     2401 gcatcaagat cagcctgata ttcttggaag gagtccagct agtgaggcag catgttcaaa
     2461 agtacccgat acgaatgtct ccttagaaga tgtgagtgaa gttgctcctg aaaagcctat
     2521 tactactgag aaccccaaac ttccttcaac agtgtctcca aatgttttta atgagacaga
     2581 attctcatta aatgtgacaa catctgccta tttggagtca ttacatggga aaaatgttaa
     2641 acatatagat gattcctccc cagaggacct gatagcagcc tttacagaaa ccagagataa
     2701 aggaatagta gatagtgaaa gaaatgcttt taaagcaata tcagagaaga tgacagactt
     2761 taaaacaact cctcctgtag aagtcttaca tgaaaatgag tccggtggtt ctgaaattaa
     2821 agacattgga agcaaataca gtgaacaaag caaagaaaca aatggaagtg agcctctagg
     2881 tgttttccct acccaaggta ctccagtagc atctcttgac ttagaacaag aacagctcac
     2941 aattaaggct cttaaagaat taggtgaaag acaggttgag aagtcaactt ctgcacagcg
     3001 tgacgcagaa ttgccttctg aagaagtact gaagcaaact ttcacatttg ctccagaatc
     3061 ttggccacag agatcatatg acatcctaga acgtaatgtc aagaatggat ctgatcttgg
     3121 gatttcccag aagcccatca ctatcagaga aactactagg gtagatgctg tttccagcct
     3181 tagcaagact gaattggtaa aaaagcatgt cctagcaaga cttctgacag acttctcagt
     3241 gcacgatctg attttctgga gagatgtgaa gaagactggg tttgtctttg gcaccacgct
     3301 gatcatgctg ctttccctgg cagctttcag tgtcatcagt gtggtttctt acctcatcct
     3361 ggctcttctc tctgtcacca tcagcttcag gatctacaag tccgtcatcc aagctgtaca
     3421 gaagtcagaa gaaggccatc cattcaaagc ctacctggac gtagacatta ctctgtcctc
     3481 agaagctttc cataattaca tgaatgctgc catggtgcac atcaacaggg ccctgaaact
     3541 cattattcgt ctctttctgg tagaagatct ggttgactcc ttgaagctgg ctgtcttcat
     3601 gtggctgatg acctatgttg gtgctgtttt taacggaatc acccttctaa ttcttgctga
     3661 actgctcatt ttcagtgtcc cgattgtcta tgagaagtac aagacccaga ttgatcacta
     3721 tgttggcatc gcccgagatc agaccaagtc aattgttgaa aagatccaag caaaactccc
     3781 tggaatcgcc aaaaaaaagg cagaaacggt accgcgggcc cgggatccac cggtcatgga
     3841 tagcaccgag gcagtgatca aggagttcat gcggttcaag gtgcacatgg agggctccat
     3901 gaacggccac gagttcgaga tcgagggcga gggcgagggc cgcccctacg agggcaccca
     3961 gaccgccaag ctgagggtga ccaagggtgg ccccctgccc ttctcctggg acatcctgtc
     4021 ccctcagttc atgtacggct ccagggcctt cacgaagcac cccgccgaca tccccgacta
     4081 ctggaagcag tccttccccg agggcttcaa gtgggagcgc gtgatgaact tcgaggacgg
     4141 cggcgccgtg tccgtggccc aggacacctc cctggaggac ggcaccctga tctacaaggt
     4201 gaagctccgc ggcaccaact tccctcctga cggccccgta atgcagaaga agacaatggg
     4261 ctgggaagca tccaccgagc ggttgtaccc cgaggacgtc gtgctgaagg gcgacattaa
     4321 gatggccctg cgcctgaagg acggcggccg ctacctggcg gacttcaaga ccacctacag
     4381 ggccaagaag cccgtgcaga tgcccggcgc cttcaacatc gaccgcaagt tggacatcac
     4441 atcccacaac gaggactaca ccgtggtgga acagtacgaa cgctccgtgg cccgccactc
     4501 caccggtggc tccggtggct cctgagcggc cgcgactcta gatcataatc agccatacca
     4561 catttgtaga ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac
     4621 ataaaatgaa tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat
     4681 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     4741 gtttgtccaa actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta
     4801 aaattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc
     4861 aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg
     4921 aacaagagtc cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat
     4981 cagggcgatg gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc
     5041 cgtaaagcac taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag
     5101 ccggcgaacg tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg
     5161 gcaagtgtag cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta
     5221 cagggcgcgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt
     5281 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     5341 taatattgaa aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt
     5401 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5461 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5521 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5581 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5641 cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5701 gaggcctagg cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt
     5761 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     5821 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     5881 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac
     5941 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     6001 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     6061 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     6121 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     6181 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     6241 caggggctcg cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag
     6301 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     6361 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     6421 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     6481 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     6541 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     6601 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     6661 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     6721 ctagggggag gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga
     6781 cggcaataaa aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg
     6841 ttcggtccca gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg
     6901 cccgcgtttc ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg
     6961 cagccaacgt cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga
     7021 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     7081 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     7141 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     7201 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     7261 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     7321 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     7381 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     7441 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     7501 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     7561 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     7621 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     7681 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     7741 ggaaaaacgc cagcaacgcg gccttttt
//
