LOCUS       sequence-221592-        7736 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses TMEM41B tagged with GFP in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7736)
  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 7736)
  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-GFP
FEATURES             Location/Qualifiers
     source          1..7736
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    739..1096
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             1161..1577
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1507..1529
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1545..1561
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    1587..1961
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1899..1915
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(2900..2902,2903..3616)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2900..2902,2903..3613)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2942..2963)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3203..3222)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3550..3571
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3647..4213
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3663..4215
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3807..3824)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4034..4053
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4233..4832
                     /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(4233..4252)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4729..4749
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             5062..5531
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5661..5678)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5832..6420)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5912..5931)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6584..7375,7376..7444))
                     /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     7207..7226
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7445..7549)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7617..7635
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg ctagagaacc atcagatgtt tccagggtgc cccaaggacc
      421 tgaaatgacc ctgtgcctta tttgaactaa ccaatcagtt cgcttctcgc ttctgttcgc
      481 gcgcttctgc tccccgagct caataaaaga gcccacaacc cctcactcgg cgcgccagtc
      541 ctccgattga ctgagtcgcc cgggtacccg tgtatccaat aaaccctctt gcagttgcat
      601 ccgacttgtg gtctcgctgt tccttgggag ggtctcctct gagtgattga ctacccgtca
      661 gcgggggtct ttcatttggg ggctcgtccg ggatcgggag acccctgccc agggaccacc
      721 gacccaccac cgggaggtaa gctggccagc aacttatctg tgtctgtccg attgtctagt
      781 gtctatgact gattttatgc gcctgcgtcg gtactagtta gctaactagc tctgtatctg
      841 gcggacccgt ggtggaactg acgagttcgg aacacccggc cgcaaccctg ggagacgtcc
      901 cagggacttc gggggccgtt tttgtggccc gacctgagtc caaaaatccc gatcgttttg
      961 gactctttgg tgcacccccc ttagaggagg gatatgtggt tctggtagga gacgagaacc
     1021 taaaacagtt cccgcctccg tctgaatttt tgctttcggt ttgggaccga agccgcgccg
     1081 cgcgtcttgt ctgctgcagc atcgttctgt gttgtctctg tctgactgtg tttctgtatt
     1141 tgtctgaaaa tatgggcccg ggccagactg ttaccactcc cttaagtttg accttaggtc
     1201 actggaaaga tgtcgagcgg atcgctcaca accagtcggt agatgtcaag aagagacgtt
     1261 gggttacctt ctgctctgca gaatggccaa cctttaacgt cggatggccg cgagacggca
     1321 cctttaaccg agacctcatc acccaggtta agatcaaggt cttttcacct ggcccgcatg
     1381 gacacccaga ccaggtcccc tacatcgtga cctgggaagc cttggctttt gacccccctc
     1441 cctgggtcaa gccctttgta caccctaagc ctccgcctcc tcttcctcca tccgccccgt
     1501 ctctccccct tgaacctcct cgttcgaccc cgcctcgatc ctccctttat ccagccctca
     1561 ctccttctct aggcgccccc atatggccat atgagatctt atatggggca cccccgcccc
     1621 ttgtaaactt ccctgaccct gacatgacaa gagttactaa cagcccctct ctccaagctc
     1681 acttacaggc tctctactta gtccagcacg aagtctggag acctctggcg gcagcctacc
     1741 aagaacaact ggaccgaccg gtggtacctc acccttaccg agtcggcgac acagtgtggg
     1801 tccgccgaca ccagactaag aacctagaac ctcgctggaa aggaccttac acagtcctgc
     1861 tgaccacccc caccgccctc aaagtagacg gcatcgcagc ttggatacac gccgcccacg
     1921 tgaaggctgc cgaccccggg ggtggaccat cctctagact gccggatctg ccatcgatat
     1981 aaagcttaaa ggatccatgg cgaaaggcag agtcgccgaa cgatcgcagt tgggcgctca
     2041 ccacacgacc cccgtggggg acggggcagc ggggacgcgg ggtctcgcgg cgcctggcag
     2101 cagagaccac cagaaggaaa aatcctgggt agaagctgga tcagcaagaa tgtcactcct
     2161 tatattggtg tccattttct tatctgcagc ttttgttatg tttttggtat ataaaaattt
     2221 tcctcagctt agtgaagaag aaagagtgaa tatgaaggtt cccagagata tggatgatgc
     2281 caaggctcta ggaaaagttt tatccaaata caaggacacc ttttatgttc aagtacttgt
     2341 agcttatttt gctacatata ttttcttgca aacatttgct attccaggct ctatatttct
     2401 cagtatactc tcagggtttc tttatccctt tccactagcc ttatttcttg tttgtttgtg
     2461 ttctggactt ggtgcctctt tctgttatat gctttcctat ttagttggga gaccagttgt
     2521 atacaaatac ctaacagaga aagcagtaaa atggtcacag caggttgaac gtcatagaga
     2581 acatctcatt aactacatta tatttttgag aataacacca tttctgccta attggtttat
     2641 taatatcaca tctcctgtga taaacgtgcc attgaaagtt ttttttattg gtacttttct
     2701 aggtgtcgca cctccttctt ttgtagccat taaggcagga acaacactgt atcaacttac
     2761 aacagcagga gaagctgttt cctggaactc aatatttatt ctgatgatct tggctgttct
     2821 ttctattctg ccagccatct tccaaaaaaa actaaagcag aaatttgagg gatccgggaa
     2881 ttcgggctct ggttcaggcg tgagcaaggg cgaggagctg ttcaccgggg tggtgcccat
     2941 cctggtcgag ctggacggcg acgtaaacgg ccacaagttc agcgtgtccg gcgagggcga
     3001 gggcgatgcc acctacggca agctgaccct gaagttcatc tgcaccaccg gcaagctgcc
     3061 cgtgccctgg cccaccctcg tgaccaccct gacctacggc gtgcagtgct tcagccgcta
     3121 ccccgaccac atgaagcagc acgacttctt caagtccgcc atgcccgaag gctacgtcca
     3181 ggagcgcacc atcttcttca aggacgacgg caactacaag acccgcgccg aggtgaagtt
     3241 cgagggcgac accctggtga accgcatcga gctgaagggc atcgacttca aggaggacgg
     3301 caacatcctg gggcacaagc tggagtacaa ctacaacagc cacaacgtct atatcatggc
     3361 cgacaagcag aagaacggca tcaaggtgaa cttcaagatc cgccacaaca tcgaggacgg
     3421 cagcgtgcag ctcgccgacc actaccagca gaacaccccc atcggcgacg gccccgtgct
     3481 gctgcccgac aaccactacc tgagcaccca gtccgccctg agcaaagacc ccaacgagaa
     3541 gcgcgatcac atggtcctgc tggagttcgt gaccgccgcc gggatcactc tcggcatgga
     3601 cgagctgtac aagtaatgat ttctcgagag cggccgctac gtaaattccg cccccccccc
     3661 taacgttact ggccgaagcc gcttggaata aggccggtgt gcgtttgtct atatgttatt
     3721 ttccaccata ttgccgtctt ttggcaatgt gagggcccgg aaacctggcc ctgtcttctt
     3781 gacgagcatt cctaggggtc tttcccctct cgccaaagga atgcaaggtc tgttgaatgt
     3841 cgtgaaggaa gcagttcctc tggaagtttc ttgaagacaa acaacgtctg tagcgaccct
     3901 ttgcaggcag cggaaccccc cacctggcga caggtgcctc tgcggccaaa agccacgtgt
     3961 ataagataca cctgcaaagg cggcacaacc ccagtgccac gttgtgagtt ggatagttgt
     4021 ggaaagagtc aaatggctct cctcaagcgt attcaacaag gggctgaagg atgcccagaa
     4081 ggtaccccat tgtatgggat ctgatctggg gcctcggtgc acatgcttta catgtgttta
     4141 gtcgaggtta aaaaaacgtc taggcccccc gaaccacggg gacgtggttt tcctttgaaa
     4201 aacacgatga taatatggcc acaaccgtcg acatgaccga gtacaagccc acggtgcgcc
     4261 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     4321 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     4381 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     4441 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     4501 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     4561 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     4621 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     4681 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     4741 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     4801 cctggtgcat gacccgcaag cccggtgcct gagtcgaccg cccgccccac gacccgcagc
     4861 gcccgaccga aaggagcgca cgaccccatg gctcgacgat aaaataaaag attttattta
     4921 gtctccagaa aaagggggga atgaaagacc ccacctgtag gtttggcaag ctagcttaag
     4981 taacgccatt ttgcaaggca tggaaaaata cataactgag aatagagaag ttcagatcaa
     5041 ggtcaggaac agatggaaca gctgaatatg ggccaaacag gatatctgtg gtaagcagtt
     5101 cctgccccgg ctcagggcca agaacagatg gaacagctga atatgggcca aacaggatat
     5161 ctgtggtaag cagttcctgc cccggctcag ggccaagaac agatggtccc cagatgcggt
     5221 ccagccctca gcagtttcta gagaaccatc agatgtttcc agggtgcccc aaggacctga
     5281 aatgaccctg tgccttattt gaactaacca atcagttcgc ttctcgcttc tgttcgcgcg
     5341 cttctgctcc ccgagctcaa taaaagagcc cacaacccct cactcggggc gccagtcctc
     5401 cgattgactg agtcgcccgg gtacccgtgt atccaataaa ccctcttgca gttgcatccg
     5461 acttgtggtc tcgctgttcc ttgggagggt ctcctctgag tgattgacta cccgtcagcg
     5521 ggggtctttc acatgcagca tgtatcaaaa ttaatttggt tttttttctt aagtatttac
     5581 attaaatggc catagttgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5641 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5701 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5761 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5821 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5881 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5941 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6001 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6061 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6121 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6181 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6241 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6301 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6361 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6421 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6481 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt gcggccggcc
     6541 gcaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa tgcttaatca
     6601 gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc tgactccccg
     6661 tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct gcaatgatac
     6721 cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca gccggaaggg
     6781 ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt aattgttgcc
     6841 gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt gccattgcta
     6901 caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc ggttcccaac
     6961 gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc
     7021 ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt atggcagcac
     7081 tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact ggtgagtact
     7141 caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa
     7201 cacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt
     7261 cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg atgtaaccca
     7321 ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct gggtgagcaa
     7381 aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa tgttgaatac
     7441 tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt ctcatgagcg
     7501 gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc acatttcccc
     7561 gaaaagtgcc acctgacgtc taagaaacca ttattatcat gacattaacc tataaaaata
     7621 ggcgtatcac gaggcccttt cgtcttcact ataggagctt ggcccattgc atacgttgta
     7681 tccatatcat aatatgtaca tttatattgg ctcatgtcca acattaccgc catgtt
//
