LOCUS       sequence-224940-        7088 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TMEM41B tagged with 3xFLAG in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7088)
  AUTHORS   Morita K, Hama Y, Izume T, Tamura N, Ueno T, Yamashita Y, Sakamaki 
            Y, Mimura K, Morishita H, Shihoya W, Nureki O, Mano H, Mizushima N
  TITLE     Genome-wide CRISPR screen identifies TMEM41B as a gene required for 
            autophagosome formation.
  JOURNAL   J Cell Biol. 2018 Aug 9. pii: jcb.201804132. doi: 
            10.1083/jcb.201804132.
  PUBMED    30093494
REFERENCE   2  (bases 1 to 7088)
  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: "J Cell 
            Biol. 2018 Aug 9. pii: jcb.201804132. doi: 10.1083/jcb.201804132."
COMMENT     Sequence Label: pMRXIP TMEM41B-3xFLAG
FEATURES             Location/Qualifiers
     source          1..7088
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..417
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     347..369
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     385..401
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    427..801
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     739..755
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             1740..1805
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    1839..2405
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1855..2407
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1999..2016)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2226..2245
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             2425..3024
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2425..2444)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2921..2941
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3254..3723
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(3853..3870)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4024..4612)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4104..4123)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4776..5567,5568..5636))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5399..5418
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5637..5741)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5809..5827
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        5929..6308
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    6667..7024
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
ORIGIN
        1 ggccagactg ttaccactcc cttaagtttg accttaggtc actggaaaga tgtcgagcgg
       61 atcgctcaca accagtcggt agatgtcaag aagagacgtt gggttacctt ctgctctgca
      121 gaatggccaa cctttaacgt cggatggccg cgagacggca cctttaaccg agacctcatc
      181 acccaggtta agatcaaggt cttttcacct ggcccgcatg gacacccaga ccaggtcccc
      241 tacatcgtga cctgggaagc cttggctttt gacccccctc cctgggtcaa gccctttgta
      301 caccctaagc ctccgcctcc tcttcctcca tccgccccgt ctctccccct tgaacctcct
      361 cgttcgaccc cgcctcgatc ctccctttat ccagccctca ctccttctct aggcgccccc
      421 atatggccat atgagatctt atatggggca cccccgcccc ttgtaaactt ccctgaccct
      481 gacatgacaa gagttactaa cagcccctct ctccaagctc acttacaggc tctctactta
      541 gtccagcacg aagtctggag acctctggcg gcagcctacc aagaacaact ggaccgaccg
      601 gtggtacctc acccttaccg agtcggcgac acagtgtggg tccgccgaca ccagactaag
      661 aacctagaac ctcgctggaa aggaccttac acagtcctgc tgaccacccc caccgccctc
      721 aaagtagacg gcatcgcagc ttggatacac gccgcccacg tgaaggctgc cgaccccggg
      781 ggtggaccat cctctagact gccggatctg ccatcgatat aaagcttaaa ggatccatgg
      841 cgaaaggcag agtcgccgaa cgatcgcagt tgggcgctca ccacacgacc cccgtggggg
      901 acggggcagc ggggacgcgg ggtctcgcgg cgcctggcag cagagaccac cagaaggaaa
      961 aatcctgggt agaagctgga tcagcaagaa tgtcactcct tatattggtg tccattttct
     1021 tatctgcagc ttttgttatg tttttggtat ataaaaattt tcctcagctt agtgaagaag
     1081 aaagagtgaa tatgaaggtt cccagagata tggatgatgc caaggctcta ggaaaagttt
     1141 tatccaaata caaggacacc ttttatgttc aagtacttgt agcttatttt gctacatata
     1201 ttttcttgca aacatttgct attccaggct ctatatttct cagtatactc tcagggtttc
     1261 tttatccctt tccactagcc ttatttcttg tttgtttgtg ttctggactt ggtgcctctt
     1321 tctgttatat gctttcctat ttagttggga gaccagttgt atacaaatac ctaacagaga
     1381 aagcagtaaa atggtcacag caggttgaac gtcatagaga acatctcatt aactacatta
     1441 tatttttgag aataacacca tttctgccta attggtttat taatatcaca tctcctgtga
     1501 taaacgtgcc attgaaagtt ttttttattg gtacttttct aggtgtcgca cctccttctt
     1561 ttgtagccat taaggcagga acaacactgt atcaacttac aacagcagga gaagctgttt
     1621 cctggaactc aatatttatt ctgatgatct tggctgttct ttctattctg ccagccatct
     1681 tccaaaaaaa actaaagcag aaatttgagg gatccgggaa ttcgggctct ggttcaggcg
     1741 actacaaaga ccatgacggt gattataaag atcatgacat cgactacaag gatgacgatg
     1801 acaagtaatg atttctcgag agcggccgct acgtaaattc cgcccccccc cctaacgtta
     1861 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     1921 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     1981 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     2041 aagcagttcc tctggaagtt tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     2101 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     2161 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     2221 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     2281 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     2341 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     2401 gataatatgg ccacaaccgt cgacatgacc gagtacaagc ccacggtgcg cctcgccacc
     2461 cgcgacgacg tcccccgggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     2521 acgcgccaca ccgtcgaccc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     2581 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     2641 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     2701 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     2761 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     2821 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     2881 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     2941 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     3001 atgacccgca agcccggtgc ctgagtcgac cgcccgcccc acgacccgca gcgcccgacc
     3061 gaaaggagcg cacgacccca tggctcgacg ataaaataaa agattttatt tagtctccag
     3121 aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagctta agtaacgcca
     3181 ttttgcaagg catggaaaaa tacataactg agaatagaga agttcagatc aaggtcagga
     3241 acagatggaa cagctgaata tgggccaaac aggatatctg tggtaagcag ttcctgcccc
     3301 ggctcagggc caagaacaga tggaacagct gaatatgggc caaacaggat atctgtggta
     3361 agcagttcct gccccggctc agggccaaga acagatggtc cccagatgcg gtccagccct
     3421 cagcagtttc tagagaacca tcagatgttt ccagggtgcc ccaaggacct gaaatgaccc
     3481 tgtgccttat ttgaactaac caatcagttc gcttctcgct tctgttcgcg cgcttctgct
     3541 ccccgagctc aataaaagag cccacaaccc ctcactcggg gcgccagtcc tccgattgac
     3601 tgagtcgccc gggtacccgt gtatccaata aaccctcttg cagttgcatc cgacttgtgg
     3661 tctcgctgtt ccttgggagg gtctcctctg agtgattgac tacccgtcag cgggggtctt
     3721 tcacatgcag catgtatcaa aattaatttg gttttttttc ttaagtattt acattaaatg
     3781 gccatagttg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     3841 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     3901 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     3961 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     4021 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     4081 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     4141 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     4201 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     4261 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     4321 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     4381 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     4441 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     4501 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     4561 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     4621 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     4681 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttgcggccgg ccgcaaatca
     4741 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4801 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4861 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4921 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4981 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5041 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     5101 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     5161 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     5221 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5281 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5341 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aacacgggat
     5401 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5461 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5521 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5581 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5641 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5701 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5761 ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc
     5821 acgaggccct ttcgtcttca ctataggagc ttggcccatt gcatacgttg tatccatatc
     5881 ataatatgta catttatatt ggctcatgtc caacattacc gccatgttga cattgattat
     5941 tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt
     6001 tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc
     6061 cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac
     6121 gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata
     6181 tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc
     6241 agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta
     6301 ttaccatgct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     6361 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     6421 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgattgact
     6481 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     6541 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     6601 catttggggg ctcgtccggg atcgggagac ccctgcccag ggaccaccga cccaccaccg
     6661 ggaggtaagc tggccagcaa cttatctgtg tctgtccgat tgtctagtgt ctatgactga
     6721 ttttatgcgc ctgcgtcggt actagttagc taactagctc tgtatctggc ggacccgtgg
     6781 tggaactgac gagttcggaa cacccggccg caaccctggg agacgtccca gggacttcgg
     6841 gggccgtttt tgtggcccga cctgagtcca aaaatcccga tcgttttgga ctctttggtg
     6901 cacccccctt agaggaggga tatgtggttc tggtaggaga cgagaaccta aaacagttcc
     6961 cgcctccgtc tgaatttttg ctttcggttt gggaccgaag ccgcgccgcg cgtcttgtct
     7021 gctgcagcat cgttctgtgt tgtctctgtc tgactgtgtt tctgtatttg tctgaaaata
     7081 tgggcccg
//
