LOCUS       sequence-443766-        5373 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of fluorescent N-terminally tagged shRNA 
            resistant Sec22b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5373)
  AUTHORS   Criado Santos N, Bouvet S, Cruz Cobo M, Mandavit M, Bermont F, 
            Castelbou C, Mansour F, Azam M, Giordano F, Nunes-Hasler P
  TITLE     Sec22b regulates phagosome maturation by promoting ORP8-mediated 
            lipid exchange at endoplasmic reticulum-phagosome contact sites.
  JOURNAL   Commun Biol. 2023 Oct 4;6(1):1008. doi: 10.1038/s42003-023-05382-0.
  PUBMED    37794132
REFERENCE   2  (bases 1 to 5373)
  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: "Commun 
            Biol."; date: "2023-10-4"; volume: "6(1):1008. doi"; pages: " 
            10.1038/s42003-023-05382-0"
COMMENT     Sequence Label: pEGFP-Sec22b-shR
FEATURES             Location/Qualifiers
     source          1..5373
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(80..99)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     690..709
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(798..1155)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      812..947
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        964..1155
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     1112..1132
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        complement(1157..1261)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      1288..1743
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1375..1394)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1585..1606
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     polyA_signal    1750..1871
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1761..1780)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1815..1834
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(2703..3413,3414..3416,3417..3419))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             complement(join(2703..3413,3414..3416,3417..3419))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2745..2766)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     3094..3113
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3353..3374
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      3416..3425
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(3464..3667)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(3493..3513)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3668..3971
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     rep_origin      complement(4146..4734)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4226..4245)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    5063..5110
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     5135..5154
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(join(5342..5373,1..763))
                     /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(join(5342..5373,1..763))
                     /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"
ORIGIN
        1 cgatgcgctg cgaatcggga gcggcgatac cgtaaagcac gaggaagcgg tcagcccatt
       61 cgccgccaag ctcttcagca atatcacggg tagccaacgc tatgtcctga tagcggtccg
      121 ccacacccag ccggccacag tcgatgaatc cagaaaagcg gccattttcc accatgatat
      181 tcggcaagca ggcatcgcca tgggtcacga cgagatcctc gccgtcgggc atgctcgcct
      241 tgagcctggc gaacagttcg gctggcgcga gcccctgatg ctcttcgtcc agatcatcct
      301 gatcgacaag accggcttcc atccgagtac gtgctcgctc gatgcgatgt ttcgcttggt
      361 ggtcgaatgg gcaggtagcc ggatcaagcg tatgcagccg ccgcattgca tcagccatga
      421 tggatacttt ctcggcagga gcaaggtgag atgacaggag atcctgcccc ggcacttcgc
      481 ccaatagcag ccagtccctt cccgcttcag tgacaacgtc gagcacagct gcgcaaggaa
      541 cgcccgtcgt ggccagccac gatagccgcg ctgcctcgtc ttgcagttca ttcagggcac
      601 cggacaggtc ggtcttgaca aaaagaaccg ggcgcccctg cgctgacagc cggaacacgg
      661 cggcatcaga gcagccgatt gtctgttgtg cccagtcata gccgaatagc ctctccaccc
      721 aagcggccgg agaacctgcg tgcaatccat cttgttcaat catgcgaaac gatcctcatc
      781 ctgtctcttg atcgatcttt gcaaaagcct aggcctccaa aaaagcctcc tcactacttc
      841 tggaatagct cagaggccga ggcggcctcg gcctctgcat aaataaaaaa aattagtcag
      901 ccatggggcg gagaatgggc ggaactgggc ggagttaggg gcgggatggg cggagttagg
      961 ggcgggacta tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
     1021 gagcctgggg actttccaca cctggttgct gactaattga gatgcatgct ttgcatactt
     1081 ctgcctgctg gggagcctgg ggactttcca caccctaact gacacacatt ccacagctgg
     1141 ttctttccgc ctcaggactc ttcctttttc aatattattg aagcatttat cagggttatt
     1201 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     1261 gcacatttcc ccgaaaagtg ccacctgacg cgccctgtag cggcgcatta agcgcggcgg
     1321 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     1381 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     1441 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     1501 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     1561 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     1621 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     1681 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     1741 tttacgcgtt aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     1801 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     1861 ataaacaagt taacaacaac aattgcattc attttatgtt tcaggttcag ggggaggtgt
     1921 gggaggtttt ttaaagcaag taaaacctct acaaatgtgg tatggctgat tatgatcagt
     1981 tatctagatc cggtggatcc cgggcccgcg gtacctcaca gccaccaaaa ccgcacatac
     2041 actatcaaca tgatgaagaa cacggctact gctgcaagct tggcgtaggt ggaacgcatg
     2101 ttcaggtact tcgcatcctg gcggtatttc ttagatagac tggacaaatt gttagccttc
     2161 gaatccaatg ctgagagtgc ttctccccgc tgtaggactt cttcaatatt cgccaccatg
     2221 atcctctgca catcttgcaa ttcggtgttg atggagccga gatttctccg agcacggcta
     2281 tcaatgtaga gtttcttggt cttctgaatg aaggtgtcga actcgataaa agaataaggc
     2341 ctagacacag tgggcacctt cttcccatgc tgctcgtcaa attccgagtg caagtcttct
     2401 aggtaggcaa aggccaactt cttagggaag gcagcttcac acaaaaccaa gtaacacacc
     2461 ccctgctcaa taatgtagtg aaaagtcatg gctcctgctt ccaaggtaca tctggtaggg
     2521 gactgttcat tcaactttcg aaagagttgc ttagcctgac tctgatactg ctgaaggtcc
     2581 cggcctgact gctcgtcctc ctgcatcgag gcggccagag gaagcccgtc cgctacgcgg
     2641 gcaatcattg tcagcagcac ggatccggaa ttcgaagctt gagctcgaga tctgagtccg
     2701 gacttgtaca gctcgtccat gccgagagtg atcccggcgg cggtcacgaa ctccagcagg
     2761 accatgtgat cgcgcttctc gttggggtct ttgctcaggg cggactgggt gctcaggtag
     2821 tggttgtcgg gcagcagcac ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg
     2881 gcgagctgca cgctgccgtc ctcgatgttg tggcggatct tgaagttcac cttgatgccg
     2941 ttcttctgct tgtcggccat gatatagacg ttgtggctgt tgtagttgta ctccagcttg
     3001 tgccccagga tgttgccgtc ctccttgaag tcgatgccct tcagctcgat gcggttcacc
     3061 agggtgtcgc cctcgaactt cacctcggcg cgggtcttgt agttgccgtc gtccttgaag
     3121 aagatggtgc gctcctggac gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc
     3181 ttcatgtggt cggggtagcg gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg
     3241 gtgggccagg gcacgggcag cttgccggtg gtgcagatga acttcagggt cagcttgccg
     3301 taggtggcat cgccctcgcc ctcgccggac acgctgaact tgtggccgtt tacgtcgccg
     3361 tccagctcga ccaggatggg caccaccccg gtgaacagct cctcgccctt gctcaccatg
     3421 gtggcgaccg gtagcgctag cggatctgac ggttcactaa accagctctg cttatataga
     3481 cctcccaccg tacacgccta ccgcccattt gcgtcaatgg ggcggagttg ttacgacatt
     3541 ttggaaagtc ccgttgattt tggtgccaaa acaaactccc attgacgtca atggggtgga
     3601 gacttggaaa tccccgtgag tcaaaccgct atccacgccc attgatgtac tgccaaaacc
     3661 gcatcaccat ggtaatagcg atgactaata cgtagatgta ctgccaagta ggaaagtccc
     3721 ataaggtcat gtactgggca taatgccagg cgggccattt accgtcattg acgtcaatag
     3781 ggggcgtact tggcatatga tacacttgat gtactgccaa gtgggcagtt taccgtaaat
     3841 actccaccca ttgacgtcaa tggaaagtcc ctattggcgt tactatggga acatacgtca
     3901 ttattgacgt caatgggcgg gggtcgttgg gcggtcagcc aggcgggcca tttaccgtaa
     3961 gttatgtaac gcggaactcc atatatgggc tatgaactaa tgaccccgta attgattact
     4021 attaataact aatgcatggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4081 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4141 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4201 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4261 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4321 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4381 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4441 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4501 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4561 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4621 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4681 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4741 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4801 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4861 tttaaatcaa tctaaagtat atatgagtaa cctgaggcta tggcagggcc tgccgccccg
     4921 acgttggctg cgagccctgg gccttcaccc gaacttgggg ggtggggtgg ggaaaaggaa
     4981 gaaacgcggg cgtattggcc ccaatggggt ctcggtgggg tatcgacaga gtgccagccc
     5041 tgggaccgaa ccccgcgttt atgaacaaac gacccaacac cgtgcgtttt attctgtctt
     5101 tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt ttcagttagc
     5161 ctccccctag ggtgggcgaa gaactccagc atgagatccc cgcgctggag gatcatccag
     5221 ccggcgtccc ggaaaacgat tccgaagccc aacctttcat agaaggcggc ggtggaatcg
     5281 aaatctcgtg atggcaggtt gggcgtcgct tggtcggtca tttcgaaccc cagagtcccg
     5341 ctcagaagaa ctcgtcaaga aggcgataga agg
//
