LOCUS       sequence-334695-        5304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression of Sar1b tagged with mOrange2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5304)
  AUTHORS   Weigel AV, Chang CL, Shtengel G, Xu CS, Hoffman DP, Freeman M, Iyer 
            N, Aaron J, Khuon S, Bogovic J, Qiu W, Hess HF, Lippincott-Schwartz 
            J
  TITLE     ER-to-Golgi protein delivery through an interwoven, tubular network 
            extending from ER.
  JOURNAL   Cell. 2021 Apr 29;184(9):2412-2429.e16. doi: 
            10.1016/j.cell.2021.03.035. Epub 2021 Apr 13.
  PUBMED    33852913
REFERENCE   2  (bases 1 to 5304)
  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.2021.03.035"; journalName: "Cell"; date: 
            "2021-04-29- 29"; volume: "184"; issue: "9"; pages: "2412-2429.e16"
COMMENT     Sequence Label: Sar1b-mOrange2
FEATURES             Location/Qualifiers
     source          1..5304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      7..142
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        159..350
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     307..327
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(352..456)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      483..938
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(570..589)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     780..801
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    945..1066
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(956..975)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1010..1029
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(1189..1899)
                     /codon_start=1
                     /product="photostable monomeric orange derivative of DsRed 
                     fluorescent protein (Shaner et al., 2008)"
                     /label=mOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPHFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GKIKMRLKLKDGGHYTSEVKTTYKAKKPVQLPGAYIVDIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     regulatory      1896..1905
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(2590..2793)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2619..2639)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2794..3097
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(3272..3860)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3352..3371)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    4189..4236
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     4261..4280
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(4468..5262)
                     /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(4468..5262)
                     /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(4579..4598)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     5189..5208
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(join(5297..5304,1..350))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
ORIGIN
        1 agcctaggcc tccaaaaaag cctcctcact acttctggaa tagctcagag gccgaggcgg
       61 cctcggcctc tgcataaata aaaaaaatta gtcagccatg gggcggagaa tgggcggaac
      121 tgggcggagt taggggcggg atgggcggag ttaggggcgg gactatggtt gctgactaat
      181 tgagatgcat gctttgcata cttctgcctg ctggggagcc tggggacttt ccacacctgg
      241 ttgctgacta attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact
      301 ttccacaccc taactgacac acattccaca gctggttctt tccgcctcag gactcttcct
      361 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
      421 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
      481 tgacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac
      541 cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc
      601 cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt
      661 tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg
      721 gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag
      781 tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt
      841 ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt
      901 taacgcgaat tttaacaaaa tattaacgct tacaatttac gccttaagat acattgatga
      961 gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga
     1021 tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg
     1081 cattcatttt atgtttcagg ttcaggggga ggtgtgggag gttttttaaa gcaagtaaaa
     1141 cctctacaaa tgtggtatgg ctgattatga tctagagtcg cggccgcttt acttgtacag
     1201 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
     1261 gtagtcctcg ttgtgggagg tgatgtccaa cttgatgtcg acgatgtagg cgccgggcag
     1321 ctgcacgggc ttcttggcct tgtaggtggt cttgacctcg gaggtgtagt ggccgccgtc
     1381 cttcagcttc agcctcatct tgatcttgcc cttcagggca ccgtcctcgg ggtacatccg
     1441 ctcggaggag gcctcccagc ccatggtctt cttctgcatc acggggccgt cggaggggaa
     1501 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     1561 ggtcacggtc accacgccgc cgtcctcgta gttcatcacg cgctcccact tgaagccctc
     1621 ggggaaggac agcttgaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     1681 gccgtaggtg aaatgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     1741 caccttcagc ttagcggtct gaaagccctc gtaggggcgg ccctcgccct cgccctcgat
     1801 ctcgaactcg tggccgttca cggagccctc catgcgcacc ttgaagcgca tgaactcctt
     1861 gatgatggcc atgttattct cctcgccctt gctcaccatg gtggcgaccg gtggatcccc
     1921 atcgatgtac tgtgccatcc agcggaagcc ttctccatag ccttgccttt tcagcacact
     1981 gcacatgaac acttccagag gtctggcatt cagctccttc agtgacacac tgccctttcc
     2041 tgttgtctgc ccatataagc caaacatctc tcgtaacctc tcttcactga tggcttcggg
     2101 tctgtcaatc ttatttccaa gaattaatat aggcacattg gcaatggttt catctgtcat
     2161 tagtgaatca agttcttctt ttgattctaa cagcctttca tggtctgcac aatccacaag
     2221 aaatacaatg ccattgatag caggaaggta atttttccat actcttcggg cttggatgtg
     2281 cccacccaga tcaaaagttg taaaagtcat gccagcaata gtgagttctt ctgaagtggg
     2341 atgtagtgtt gggacgtgct ggccaagtct gtcatctttc agcatgtgta gcaacgtggt
     2401 tttcccagca ttgtccaatc caaggaatac cagtttacca gtttttttat acagtcctag
     2461 aaactgcagc acactgctga aaccactgta aatccagtca aatatgaagg acatggtggc
     2521 ggtgaattcg aagcttgagc tcgagatctg agtccggtag cgctagcgga tctgacggtt
     2581 cactaaacca gctctgctta tatagacctc ccaccgtaca cgcctaccgc ccatttgcgt
     2641 caatggggcg gagttgttac gacattttgg aaagtcccgt tgattttggt gccaaaacaa
     2701 actcccattg acgtcaatgg ggtggagact tggaaatccc cgtgagtcaa accgctatcc
     2761 acgcccattg atgtactgcc aaaaccgcat caccatggta atagcgatga ctaatacgta
     2821 gatgtactgc caagtaggaa agtcccataa ggtcatgtac tgggcataat gccaggcggg
     2881 ccatttaccg tcattgacgt caataggggg cgtacttggc atatgataca cttgatgtac
     2941 tgccaagtgg gcagtttacc gtaaatactc cacccattga cgtcaatgga aagtccctat
     3001 tggcgttact atgggaacat acgtcattat tgacgtcaat gggcgggggt cgttgggcgg
     3061 tcagccaggc gggccattta ccgtaagtta tgtaacgcgg aactccatat atgggctatt
     3121 aactaatgac cccgtaattg attactatta ataactaatg catggcggta atacggttat
     3181 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     3241 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     3301 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     3361 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     3421 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     3481 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     3541 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     3601 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     3661 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     3721 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3781 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3841 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3901 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3961 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaacctg
     4021 aggctatggc agggcctgcc gccccgacgt tggctgcgag ccctgggcct tcacccgaac
     4081 ttggggggtg gggtggggaa aaggaagaaa cgcgggcgta ttggccccaa tggggtctcg
     4141 gtggggtatc gacagagtgc cagccctggg accgaacccc gcgtttatga acaaacgacc
     4201 caacaccgtg cgttttattc tgtcttttta ttgccgtcat agcgcgggtt ccttccggta
     4261 ttgtctcctt ccgtgtttca gttagcctcc ccctagggtg ggcgaagaac tccagcatga
     4321 gatccccgcg ctggaggatc atccagccgg cgtcccggaa aacgattccg aagcccaacc
     4381 tttcatagaa ggcggcggtg gaatcgaaat ctcgtgatgg caggttgggc gtcgcttggt
     4441 cggtcatttc gaaccccaga gtcccgctca gaagaactcg tcaagaaggc gatagaaggc
     4501 gatgcgctgc gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc
     4561 gccgccaagc tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc
     4621 cacacccagc cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt
     4681 cggcaagcag gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgctcgcctt
     4741 gagcctggcg aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg
     4801 atcgacaaga ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg
     4861 gtcgaatggg caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat
     4921 ggatactttc tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc
     4981 caatagcagc cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac
     5041 gcccgtcgtg gccagccacg atagccgcgc tgcctcgtct tgcagttcat tcagggcacc
     5101 ggacaggtcg gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc
     5161 ggcatcagag cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca
     5221 agcggccgga gaacctgcgt gcaatccatc ttgttcaatc atgcgaaacg atcctcatcc
     5281 tgtctcttga tcgatctttg caaa
//
