LOCUS       sequence-413177-        4911 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of NbGFP-miRFP670nano3 localized to endoplasmic
            reticulum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4911)
  AUTHORS   Sekhon H, Ha JH, Presti MF, Procopio SB, Jarvis AR, Mirsky PO, John 
            AM, Loh SN
  TITLE     Adaptable, turn-on maturation (ATOM) fluorescent biosensors for 
            multiplexed detection in cells.
  JOURNAL   Nat Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w.
  PUBMED    37945909
REFERENCE   2  (bases 1 to 4911)
  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 
            Methods. 2023 Nov 9. doi: 10.1038/s41592-023-02065-w."
COMMENT     Sequence Label: pCMV-PRLSS-NbGFP_miRFP670nano3-KDEL
FEATURES             Location/Qualifiers
     source          1..4911
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     polyA_signal    1699..1820
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1736..1755)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1790..1809
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1827..2282)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1964..1985)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2176..2195
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2309..2413
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2415..2772
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2415..2606
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2438..2458)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2623..2758
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2807..3601
                     /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             2807..3601
                     /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(2861..2880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3471..3490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3789..3808)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3833..3880
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4209..4797
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4698..4717
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attctgccac catgaacatc aaaggatcgc
      661 catggaaagg gtccctcctg ctgctgctgg tgtcaaacct gctcctgtgc cagagcgtgg
      721 caccggtctc aggtgctcaa gtccaactgg tggagtctgg tggcgctttg gtgcagccag
      781 gtggctctct gcgtttgtcc tgtgccgctt ctggcttccc agtgaaccgc tattccatgc
      841 gctggtatcg ccaggctcca ggcaaagagc gtgagtgggt agccggtatg tccagcgcgg
      901 gtgatcgtag ctcctatgaa gactccggcg gagggggtag catggcaaac ctggacaaga
      961 tgctgaacac caccgtgacc gaggtgcgca agttcctgca agcggacaga gtgtgcgtgt
     1021 tcaagttcga ggaagattac tccggcaccg tctcgcacga agccgtggac gacagatgga
     1081 ttagcatcct gaagacccag gtgcaggaca gatacttcat ggaaaccaga ggcgaggaat
     1141 acgtccacgg cagataccag gccatcgccg acatctacac agccaatctg gtcgagtgct
     1201 acagagacct gctgatcgag tttcaggtgc gggccattct ggctgtcccc atcctgcaag
     1261 gcaagaagct gtggggcctg ctggtggccc accaactggc cggccctcgg gagtggcaga
     1321 cctgggaaat cgacttcctg aaacagcaag ccgtggtgat gggcatcgcc atccagcaga
     1381 gcggcggagg gggtagcgtg aagggccgtt tcaccatcag ccgtgacgat gcccgtaaca
     1441 cggtgtatct gcaaatgaac agcttgaaac ctgaagatac ggccgtgtat tactgtaatg
     1501 tgaacgtggg cttcgagtat tggggccaag gcacccaggt caccgtctcc agcggtagcg
     1561 gtaaagacga gctgtgagcg gccgcgactc tagatcataa tcagccatac cacatttgta
     1621 gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg
     1681 aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1741 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1801 aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc
     1861 gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc
     1921 ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag
     1981 tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga
     2041 tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc
     2101 actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa
     2161 cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt
     2221 agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc
     2281 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     2341 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     2401 aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt
     2461 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2521 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2581 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2641 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2701 gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2761 ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga
     2821 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     2881 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     2941 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3001 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3061 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3121 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3181 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3241 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3301 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3361 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3421 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3481 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3541 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3601 agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     3661 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     3721 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg
     3781 aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata
     3841 aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc
     3901 cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt
     3961 tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac
     4021 gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta gattgattta
     4081 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4141 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4201 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4261 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4321 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4381 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4441 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4501 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4561 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4621 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4681 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4741 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4801 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4861 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
