LOCUS       sequence-461113-        6497 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Visualization of ER in mammalian cells using mGold2t.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6497)
  AUTHORS   Lee J, Lai S, Yang S, Zhao S, Blanco FA, Lyons AC, Merino-Urteaga R,
            Ahrens JF, Nguyen NA, Liu H, Liu Z, Lambert GG, Shaner NC, Chen L, 
            Tolias KF, Zhang J, Ha T, St-Pierre F
  TITLE     Bright and photostable yellow fluorescent proteins for extended 
            imaging.
  JOURNAL   Nat Commun. 2025 Apr 4;16(1):3241. doi: 10.1038/s41467-025-58223-5.
  PUBMED    40185748
REFERENCE   2  (bases 1 to 6497)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat 
            Commun."; date: "2025-04-4"; volume: "16(1):3241. doi"; pages: " 
            10.1038/s41467-025-58223-5"
COMMENT     Sequence Label: pCMV-mGold2t-Calnexin-N-14
FEATURES             Location/Qualifiers
     source          1..6497
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        76..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2452..2461
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2458..2460,2461..2463,2464..3177)
                     /codon_start=1
                     /product="photostable derivative of mVenus (Lee et al., 
                     2020)"
                     /label=mGold
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDINGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTSLGYGLQCFARYPDNMKRHDFFKSAMPEGYVQERTIFFEDD
                     GNYKTRAEVKFEGGTLVNRVELKGIDFKEDGNILGHKLEYNYNCHNVFITADRQQNGIK
                     VNFKIRHNIEVGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQTKLSKDPNEKRDHMVLL
                     EFVTAAGITLSMDELYK"
     primer_bind     complement(2764..2783)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3111..3132
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3300..3421
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3337..3356)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3391..3410
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3428..3883)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3565..3586)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3777..3796
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3910..4014
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4016..4373
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4016..4207
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4039..4059)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4224..4359
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4408..5202
                     /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             4408..5202
                     /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(4462..4481)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5072..5091
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5390..5409)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5434..5481
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5810..6398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6299..6318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 attaccgcca tgcattagtt attaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgctaccgga ctcagatctc gagaccgcca tggaagggaa gtggttgctg
      661 tgtatgttac tggtgcttgg aactgctatt gttgaggctc atgatggaca tgatgatgat
      721 gtgattgata ttgaggatga ccttgacgat gtcattgaag aggtagaaga ctcaaaacca
      781 gataccactg ctcctccttc atctcccaag gttacttaca aagctccagt tccaacaggg
      841 gaagtatatt ttgctgattc ttttgacaga ggaactctgt cagggtggat tttatccaaa
      901 gccaagaaag acgataccga tgatgaaatt gccaaatatg atggaaagtg ggaggtagag
      961 gaaatgaagg agtcaaagct tccaggtgat aaaggacttg tgttgatgtc tcgggccaag
     1021 catcatgcca tctctgctaa actgaacaag cccttcctgt ttgacaccaa gcctctcatt
     1081 gttcagtatg aggttaattt ccaaaatgga atagaatgtg gtggtgccta tgtgaaactg
     1141 ctttctaaaa caccagaact caacctggat cagttccatg acaagacccc ttatacgatt
     1201 atgtttggtc cagataaatg tggagaggac tataaactgc acttcatctt ccgacacaaa
     1261 aaccccaaaa cgggtatcta tgaagaaaaa catgctaaga ggccagatgc agatctgaag
     1321 acctatttta ctgataagaa aacacatctt tacacactaa tcttgaatcc agataatagt
     1381 tttgaaatac tggttgacca atctgtggtg aatagtggaa atctgctcaa tgacatgact
     1441 cctcctgtaa atccttcacg tgaaattgag gacccagaag accggaagcc cgaggattgg
     1501 gatgaaagac caaaaatccc agatccagaa gctgtcaagc cagatgactg ggatgaagat
     1561 gcccctgcta agattccaga tgaagaggcc acaaaacccg aaggctggtt agatgatgag
     1621 cctgagtacg tacctgatcc agacgcagag aaacctgagg attgggatga agacatggat
     1681 ggagaatggg aggctcctca gattgccaac cctagatgtg agtcagctcc tggatgtggt
     1741 gtctggcagc gacctgtgat tgacaacccc aattataaag gcaaatggaa gcctcctatg
     1801 attgacaatc ccagttacca gggaatctgg aaacccagga aaataccaaa tccagatttc
     1861 tttgaagatc tggaaccttt cagaatgact ccttttagtg ctattggttt ggagctgtgg
     1921 tccatgacct ctgacatttt ttttgacaac tttatcattt gtgctgatcg aagaatagtt
     1981 gatgattggg ccaatgatgg atggggcctg aagaaagctg ctgatggggc tgctgagcca
     2041 ggcgttgtgg ggcagatgaa cgaggcagct gaagagcgcc cgtggctgtg ggtagtctat
     2101 attctaactg tagcccttcc tgtgttcctg gttatcctct tctgctgttc tggaaagaaa
     2161 cagaccagtg gtatggagta taagaaaact gatgcacctc aaccggatgt gaaggaagag
     2221 gaagaagaga aggaagagga aaaggacaag ggagatgagg aggaggaagg agaagagaaa
     2281 cttgaagaga aacagaaaag tgatgctgaa gaagatggtg gcactgtcag tcaagaggag
     2341 gaagacagaa aacctaaagc agaggaggat gaaattttga acagatcacc aagaaacaga
     2401 aagccacgaa gagagggggg tgcgtcaggc ggcagcggat ctggaccggt cgccaccatg
     2461 gcgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc
     2521 gacataaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc
     2581 aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc
     2641 gtgaccagcc tgggctacgg cctgcagtgc ttcgcccgct accccgacaa catgaagcgg
     2701 cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc
     2761 gaggacgacg gcaactacaa gacccgcgcc gaggtgaagt ttgagggcgg caccctggtg
     2821 aaccgcgtcg agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag
     2881 ctggagtaca actacaactg ccacaacgtc tttatcaccg ccgacaggca gcagaacggc
     2941 atcaaggtca acttcaagat ccgccacaac atcgaggtcg gcggcgtgca gctcgccgac
     3001 cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac
     3061 ctgagctacc agaccaagct gagcaaagac cccaacgaga agcgcgatca catggtcctg
     3121 ctggagttcg tgaccgccgc cgggatcact ctcagcatgg acgagctgta caagtaaagc
     3181 ggccgcgact ctagatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     3241 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     3301 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     3361 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     3421 aaggcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc
     3481 agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag
     3541 accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg
     3601 gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca
     3661 tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa
     3721 gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg
     3781 aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta
     3841 accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg cacttttcgg
     3901 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     3961 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtcctgag
     4021 gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     4081 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     4141 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4201 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     4261 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     4321 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaagatcgat
     4381 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     4441 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     4501 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     4561 gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc gtggctggcc
     4621 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     4681 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     4741 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     4801 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     4861 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     4921 gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     4981 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     5041 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     5101 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     5161 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     5221 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     5281 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     5341 gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact gaaacacgga
     5401 aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc
     5461 acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     5521 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     5581 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     5641 ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat ttttaattta
     5701 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     5761 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     5821 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     5881 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     5941 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6001 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6061 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6121 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6181 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6241 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6301 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6361 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6421 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     6481 attctgtgga taaccgt
//
