LOCUS       sequence-429909-        5429 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of fluorescent N-terminally tagged shRNA 
            resistant Sec22b-P33 mutant.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5429)
  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 5429)
  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-P33-shR
FEATURES             Location/Qualifiers
     source          1..5429
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1..20)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     611..630
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(929..948)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    973..1020
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1349..1937
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1838..1857
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2112..2415
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2416..2619
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2570..2590
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      2658..2667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2664..2666,2667..2669,2670..3380)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2664..2666,2667..2669,2670..3380)
                     /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(2709..2730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2970..2989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3317..3338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4268..4389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4305..4324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4359..4378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4396..4851)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4533..4554)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4745..4764
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4878..4982
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4984..5341
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4984..5175
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5007..5027)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5192..5327
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5376..5429,1..741)
                     /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             join(5376..5429,1..741)
                     /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 ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg
       61 ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat
      121 gaactgcaag acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca
      181 gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg
      241 gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat
      301 gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa
      361 catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg
      421 gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg
      481 cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg
      541 gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat
      601 caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac
      661 cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc
      721 cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc
      781 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
      841 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
      901 tcttcgccca ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa
      961 cccgcgctat gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc
     1021 ataaacgcgg ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt
     1081 ggggccaata cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag
     1141 gcccagggct cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat
     1201 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1261 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1321 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1381 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1441 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1501 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1561 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1621 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1681 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     1741 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     1801 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     1861 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     1921 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     1981 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2041 ccgccatgca ttagttatta atagtaatca attacggggt cattagttca tagcccatat
     2101 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
     2161 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
     2221 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
     2281 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
     2341 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
     2401 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
     2461 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
     2521 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
     2581 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
     2641 cgctagcgct accggtcgcc accatggtga gcaagggcga ggagctgttc accggggtgg
     2701 tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg
     2761 agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca
     2821 agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca
     2881 gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct
     2941 acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg
     3001 tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg
     3061 aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata
     3121 tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg
     3181 aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc
     3241 ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca
     3301 acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg
     3361 gcatggacga gctgtacaag tccggactca gatctcgagt gacaatgatt gcccgcgtag
     3421 cggacgggct tcctctggcc gcctcgatgc aggaggacga gcagtcaggc cgggaccttc
     3481 agcagtatca gagtcaggct aagcaactct ttcgaaagtt gaatgaacag tcccctacca
     3541 gatgtacctt ggaagcagga gccatgactt ttcactacat tattgagcag ggggtgtgtt
     3601 acttggtttt gtgtgaagct gccttcccta agaagttggc ctttgcctac ctagaagact
     3661 tgcactcgga atttgacgag cagcatggga agaaggtgcc cactgtgtct aggccttatt
     3721 cttttatcga gttcgacacc ttcattcaga agaccaagaa actctacatt gatagccgtg
     3781 ctcggagaaa tctcggctcc atcaacaccg aattgcaaga tgtgcagagg atcatggtgg
     3841 cgaatattga agaagtccta cagcggggag aagcactctc agcattggat tcgaaggcta
     3901 acaatttgtc cagtctatct aagaaatacc gccaggatgc gaagtacctg aacatgcgat
     3961 cgcctccccc accgccgcct ccccctccac cgcccccgcc accccctcct ccacccccgc
     4021 cgccaccacc tccgcccccg ccgccaccac ctccccctcc gcgatcgacc tacgccaagc
     4081 ttgcagcagt agccgtgttc ttcatcatgt tgatagtgta tgtgcggttt tggtggctgg
     4141 atccaccgga tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     4201 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4261 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4321 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4381 tgtatcttaa cgcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     4441 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     4501 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     4561 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     4621 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     4681 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     4741 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     4801 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     4861 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     4921 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4981 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     5041 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     5101 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     5161 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     5221 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     5281 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     5341 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     5401 caggttctcc ggccgcttgg gtggagagg
//
