LOCUS       sequence-467786-        4757 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of endoplasmic reticulum marker mRuby2-PTPb.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4757)
  AUTHORS   Jang W, Senarath K, Feinberg G, Lu S, Lambert NA
  TITLE     Visualization of endogenous G proteins on endosomes and other 
            organelles.
  JOURNAL   Elife. 2024 Nov 8;13:RP97033. doi: 10.7554/eLife.97033.
  PUBMED    39514269
REFERENCE   2  (bases 1 to 4757)
  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: "Elife."; 
            date: "2024-11-8"; volume: "13:RP97033. doi"; pages: " 
            10.7554/eLife.97033"
COMMENT     Sequence Label: mRuby2-PTP1b
FEATURES             Location/Qualifiers
     source          1..4757
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        764..1067
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1068..1271
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1222..1242
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1310..1319
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1316..2026
                     /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"
     polyA_signal    2248..2369
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2285..2304)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2339..2358
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2376..2831)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2513..2534)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2725..2744
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2858..2962
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2964..3321
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2964..3155
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2987..3007)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3172..3307
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3356..4150
                     /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             3356..4150
                     /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(3410..3429)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4020..4039
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4338..4357)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4382..4429
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccatg cattagttat taatagtaat
      721 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      781 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      841 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      901 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      961 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
     1021 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
     1081 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
     1141 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
     1201 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
     1261 taagcagagc tggtttagtg aaccgtcaga tccgctagcg ctaccggtcg ccaccatggt
     1321 gtctaagggc gaagagctga tcaaggaaaa tatgcgtatg aaggtggtca tggaaggttc
     1381 ggtcaacggc caccaattca aatgcacagg tgaaggagaa ggcaatccgt acatgggaac
     1441 tcaaaccatg aggatcaaag tcatcgaggg aggacccctg ccatttgcct ttgacattct
     1501 tgccacgtcg ttcatgtatg gcagccgtac ttttatcaag tacccgaaag gcattcctga
     1561 tttctttaaa cagtcctttc ctgagggttt tacttgggaa agagttacga gatacgaaga
     1621 tggtggagtc gtcaccgtca tgcaggacac cagccttgag gatggctgtc tcgtttacca
     1681 cgtccaagtc agaggggtaa actttccctc caatggtccc gtgatgcaga agaagaccaa
     1741 gggttgggag cctaatacag agatgatgta tccagcagat ggtggtctga ggggatacac
     1801 tcatatggca ctgaaagttg atggtggtgg ccatctgtct tgctctttcg taacaactta
     1861 caggtcaaaa aagaccgtcg ggaacatcaa gatgcccggt atccatgccg ttgatcaccg
     1921 cctggaaagg ttagaggaaa gtgacaatga aatgttcgta gtacaacgcg aacacgcagt
     1981 tgccaagttc gccgggcttg gtggtgggat ggacgagctg tacaagggtg gcggagggtt
     2041 cctggtcaac atgtgcgtgg ctacggtcct cacggccggc gcttacctct gctacaggtt
     2101 cctgttcaac agcaacacat agtaaagcgg ccgcgactct agatcataat cagccatacc
     2161 acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct gaacctgaaa
     2221 cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     2281 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     2341 ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt
     2401 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2461 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2521 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2581 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2641 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2701 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2761 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2821 acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     2881 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     2941 ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag
     3001 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     3061 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     3121 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     3181 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     3241 gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt
     3301 ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt ttcgcatgat
     3361 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3421 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3481 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcaaga
     3541 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3601 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3661 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     3721 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     3781 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     3841 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc ccgacggcga
     3901 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3961 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4021 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4081 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4141 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4201 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4261 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4321 cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac ccgcgctatg
     4381 acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca taaacgcggg
     4441 gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg gggccaatac
     4501 gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg cccagggctc
     4561 gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata tatactttag
     4621 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     4681 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     4741 aagatcaaag gatcttc
//
