LOCUS       sequence-370229-        6018 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mOrange2-TM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6018)
  AUTHORS   Shi Z, Graber ZT, Baumgart T, Stone HA, Cohen AE
  TITLE     Cell Membranes Resist Flow.
  JOURNAL   Cell. 2018 Dec 13;175(7):1769-1779.e13. doi: 
            10.1016/j.cell.2018.09.054. Epub 2018 Nov 1.
  PUBMED    30392960
REFERENCE   2  (bases 1 to 6018)
  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"; doi: 
            "10.1016/j.cell.2018.09.054"; journalName: "Cell"; date: 
            "2018-12-13- 13"; volume: "175"; issue: "7"; pages: "1769-1779.e13"
COMMENT     Sequence Label: PZ055:mOrange2-TM
FEATURES             Location/Qualifiers
     source          1..6018
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     614..633
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(852..956)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      983..1438
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1070..1089)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1280..1301
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     enhancer        1450..1829
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1830..2033
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1984..2004
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2078..2097
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2078..2096
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     2177..2236
                     /codon_start=1
                     /product="leader sequence from mouse immunoglobulin kappa 
                     light chain"
                     /label=Ig-kappa leader
                     /translation="METDTLLLWVLLLWVPGSTG"
     CDS             2240..2266
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2276..2980
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="VSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGF
                     QTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNYE
                     DGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKG
                     KIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERAE
                     GRHSTGGMDELYK"
     CDS             3008..3037
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3041..3187
                     /codon_start=1
                     /product="transmembrane domain from platelet derived growth
                     factor receptor beta"
                     /label=PDGFR-beta TM domain
                     /translation="AVGQDTQEVIVVPHSLPFKVVVISAILALVVLTIISLIILIMLWQ
                     KKPR"
     primer_bind     complement(3209..3226)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3215..3439
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(3571..4159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3651..3670)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4488..4535
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4560..4579
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4767..5561)
                     /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             complement(4767..5561)
                     /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(4878..4897)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5488..5507
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(5660..5679)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        5758..5949
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     5906..5926
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(6009..6018,1..782,783..851))
                     /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"
ORIGIN
        1 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
       61 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
      121 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
      181 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
      241 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
      301 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
      361 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
      421 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
      481 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
      541 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
      601 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
      661 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
      721 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
      781 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
      841 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
      901 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
      961 tttccccgaa aagtgccacc tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     1021 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     1081 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     1141 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     1201 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     1261 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     1321 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     1381 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttac
     1441 gcgcgcgttg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     1501 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     1561 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     1621 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     1681 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     1741 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     1801 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     1861 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     1921 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     1981 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctctctggc
     2041 taactagaga acccactgct tactggctta tcgaaattaa tacgactcac tatagggaga
     2101 cccaagcttg gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgg
     2161 cttggggata tccaccatgg agacagacac actcctgcta tgggtactgc tgctctgggt
     2221 tccaggttcc actggtgact atccatatga tgttccagat tatgctgggg cccaggtgag
     2281 caagggcgag gagaataaca tggccatcat caaggagttc atgcgcttca aggtgcgcat
     2341 ggagggctcc gtgaacggcc acgagttcga gatcgagggc gagggcgagg gccgccccta
     2401 cgagggcttt cagaccgcta agctgaaggt gaccaagggt ggccccctgc ccttcgcctg
     2461 ggacatcctg tcccctcatt tcacctacgg ctccaaggcc tacgtgaagc accccgccga
     2521 catccccgac tacttcaagc tgtccttccc cgagggcttc aagtgggagc gcgtgatgaa
     2581 ctacgaggac ggcggcgtgg tgaccgtgac ccaggactcc tccctgcagg acggcgagtt
     2641 catctacaag gtgaagctgc gcggcaccaa cttcccctcc gacggccccg tgatgcagaa
     2701 gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg gtgccctgaa
     2761 gggcaagatc aagatgaggc tgaagctgaa ggacggcggc cactacacct ccgaggtcaa
     2821 gaccacctac aaggccaaga agcccgtgca gctgcccggc gcctacatcg tcgacatcaa
     2881 gttggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg aacgcgccga
     2941 gggccgccac tccaccggcg gcatggacga gctgtacaag cggggatccc cgcggctgca
     3001 ggtcgacgaa caaaaactca tctcagaaga ggatctgaat gctgtgggcc aggacacgca
     3061 ggaggtcatc gtggtgccac actccttgcc ctttaaggtg gtggtgatct cagccatcct
     3121 ggccctggtg gtgctcacca tcatctccct tatcatcctc atcatgcttt ggcagaagaa
     3181 gccacgttag gcggccgctc gagatcagcc tcgactgtgc cttctagttg ccagccatct
     3241 gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt
     3301 tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg
     3361 ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg
     3421 gatgcggtgg gctctatggc ttctgaggcg gaaagaacca gtggcggtaa tacggttatc
     3481 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     3541 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     3601 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     3661 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     3721 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     3781 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     3841 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     3901 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     3961 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     4021 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4081 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4141 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4201 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     4261 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaacctga
     4321 ggctatggca gggcctgccg ccccgacgtt ggctgcgagc cctgggcctt cacccgaact
     4381 tggggggtgg ggtggggaaa aggaagaaac gcgggcgtat tggccccaat ggggtctcgg
     4441 tggggtatcg acagagtgcc agccctggga ccgaaccccg cgtttatgaa caaacgaccc
     4501 aacaccgtgc gttttattct gtctttttat tgccgtcata gcgcgggttc cttccggtat
     4561 tgtctccttc cgtgtttcag ttagcctccc cctagggtgg gcgaagaact ccagcatgag
     4621 atccccgcgc tggaggatca tccagccggc gtcccggaaa acgattccga agcccaacct
     4681 ttcatagaag gcggcggtgg aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc
     4741 ggtcatttcg aaccccagag tcccgctcag aagaactcgt caagaaggcg atagaaggcg
     4801 atgcgctgcg aatcgggagc ggcgataccg taaagcacga ggaagcggtc agcccattcg
     4861 ccgccaagct cttcagcaat atcacgggta gccaacgcta tgtcctgata gcggtccgcc
     4921 acacccagcc ggccacagtc gatgaatcca gaaaagcggc cattttccac catgatattc
     4981 ggcaagcagg catcgccatg ggtcacgacg agatcctcgc cgtcgggcat gctcgccttg
     5041 agcctggcga acagttcggc tggcgcgagc ccctgatgct cttcgtccag atcatcctga
     5101 tcgacaagac cggcttccat ccgagtacgt gctcgctcga tgcgatgttt cgcttggtgg
     5161 tcgaatgggc aggtagccgg atcaagcgta tgcagccgcc gcattgcatc agccatgatg
     5221 gatactttct cggcaggagc aaggtgagat gacaggagat cctgccccgg cacttcgccc
     5281 aatagcagcc agtcccttcc cgcttcagtg acaacgtcga gcacagctgc gcaaggaacg
     5341 cccgtcgtgg ccagccacga tagccgcgct gcctcgtctt gcagttcatt cagggcaccg
     5401 gacaggtcgg tcttgacaaa aagaaccggg cgcccctgcg ctgacagccg gaacacggcg
     5461 gcatcagagc agccgattgt ctgttgtgcc cagtcatagc cgaatagcct ctccacccaa
     5521 gcggccggag aacctgcgtg caatccatct tgttcaatca tgcgaaacga tcctcatcct
     5581 gtctcttgat cgatctttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg
     5641 gaatagctca gaggccgggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     5701 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     5761 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     5821 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     5881 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     5941 ccgcctcagg actcttcctt tttcaataaa tcaatctaaa gtatatatga gtaaacttgg
     6001 tctgacagtt accaatgc
//
