LOCUS       sequence-224939-        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"
     misc_feature    145..502
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             567..983
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     913..935
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     951..967
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     misc_feature    993..1367
                     /label=pol region
                     /note="Moloney murine leukemia virus (MMLV) pol region 
                     containing the splice acceptor site"
     primer_bind     1305..1321
                     /label=pBMN 5'
                     /note="MMLV sequence, for inserts in pBMN retroviral 
                     vector"
     CDS             join(2306..2308,2309..3022)
                     /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(2306..2308,2309..3019)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2348..2369)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2609..2628)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2956..2977
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    3053..3619
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    3069..3621
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3213..3230)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3440..3459
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3639..4238
                     /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(3639..3658)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4135..4155
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             4468..4937
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5067..5084)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5238..5826)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5318..5337)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5990..6781,6782..6850))
                     /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     6613..6632
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6851..6955)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7023..7041
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     enhancer        7143..7522
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttgcatccga cttgtggtct cgctgttcct tgggagggtc tcctctgagt gattgactac
       61 ccgtcagcgg gggtctttca tttgggggct cgtccgggat cgggagaccc ctgcccaggg
      121 accaccgacc caccaccggg aggtaagctg gccagcaact tatctgtgtc tgtccgattg
      181 tctagtgtct atgactgatt ttatgcgcct gcgtcggtac tagttagcta actagctctg
      241 tatctggcgg acccgtggtg gaactgacga gttcggaaca cccggccgca accctgggag
      301 acgtcccagg gacttcgggg gccgtttttg tggcccgacc tgagtccaaa aatcccgatc
      361 gttttggact ctttggtgca ccccccttag aggagggata tgtggttctg gtaggagacg
      421 agaacctaaa acagttcccg cctccgtctg aatttttgct ttcggtttgg gaccgaagcc
      481 gcgccgcgcg tcttgtctgc tgcagcatcg ttctgtgttg tctctgtctg actgtgtttc
      541 tgtatttgtc tgaaaatatg ggcccgggcc agactgttac cactccctta agtttgacct
      601 taggtcactg gaaagatgtc gagcggatcg ctcacaacca gtcggtagat gtcaagaaga
      661 gacgttgggt taccttctgc tctgcagaat ggccaacctt taacgtcgga tggccgcgag
      721 acggcacctt taaccgagac ctcatcaccc aggttaagat caaggtcttt tcacctggcc
      781 cgcatggaca cccagaccag gtcccctaca tcgtgacctg ggaagccttg gcttttgacc
      841 cccctccctg ggtcaagccc tttgtacacc ctaagcctcc gcctcctctt cctccatccg
      901 ccccgtctct cccccttgaa cctcctcgtt cgaccccgcc tcgatcctcc ctttatccag
      961 ccctcactcc ttctctaggc gcccccatat ggccatatga gatcttatat ggggcacccc
     1021 cgccccttgt aaacttccct gaccctgaca tgacaagagt tactaacagc ccctctctcc
     1081 aagctcactt acaggctctc tacttagtcc agcacgaagt ctggagacct ctggcggcag
     1141 cctaccaaga acaactggac cgaccggtgg tacctcaccc ttaccgagtc ggcgacacag
     1201 tgtgggtccg ccgacaccag actaagaacc tagaacctcg ctggaaagga ccttacacag
     1261 tcctgctgac cacccccacc gccctcaaag tagacggcat cgcagcttgg atacacgccg
     1321 cccacgtgaa ggctgccgac cccgggggtg gaccatcctc tagactgccg gatctgccat
     1381 cgatataaag cttaaaggat ccatggcgaa aggcagagtc gccgaacgat cgcagttggg
     1441 cgctcaccac acgacccccg tgggggacgg ggcagcgggg acgcggggtc tcgcggcgcc
     1501 tggcagcaga gaccaccaga aggaaaaatc ctgggtagaa gctggatcag caagaatgtc
     1561 actccttata ttggtgtcca ttttcttatc tgcagctttt gttatgtttt tggtatataa
     1621 aaattttcct cagcttagtg aagaagaaag agtgaatatg aaggttccca gagatatgga
     1681 tgatgccaag gctctaggaa aagttttatc caaatacaag gacacctttt atgttcaagt
     1741 acttgtagct tattttgcta catatatttt cttgcaaaca tttgctattc caggctctat
     1801 atttctcagt atactctcag ggtttcttta tccctttcca ctagccttat ttcttgtttg
     1861 tttgtgttct ggacttggtg cctctttctg ttatatgctt tcctatttag ttgggagacc
     1921 agttgtatac aaatacctaa cagagaaagc agtaaaatgg tcacagcagg ttgaacgtca
     1981 tagagaacat ctcattaact acattatatt tttgagaata acaccatttc tgcctaattg
     2041 gtttattaat atcacatctc ctgtgataaa cgtgccattg aaagtttttt ttattggtac
     2101 ttttctaggt gtcgcacctc cttcttttgt agccattaag gcaggaacaa cactgtatca
     2161 acttacaaca gcaggagaag ctgtttcctg gaactcaata tttattctga tgatcttggc
     2221 tgttctttct attctgccag ccatcttcca aaaaaaacta aagcagaaat ttgagggatc
     2281 cgggaattcg ggctctggtt caggcgtgag caagggcgag gagctgttca ccggggtggt
     2341 gcccatcctg gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga
     2401 gggcgagggc gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa
     2461 gctgcccgtg ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag
     2521 ccgctacccc gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta
     2581 cgtccaggag cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt
     2641 gaagttcgag ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga
     2701 ggacggcaac atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat
     2761 catggccgac aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga
     2821 ggacggcagc gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc
     2881 cgtgctgctg cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa
     2941 cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg
     3001 catggacgag ctgtacaagt aatgatttct cgagagcggc cgctacgtaa attccgcccc
     3061 cccccctaac gttactggcc gaagccgctt ggaataaggc cggtgtgcgt ttgtctatat
     3121 gttattttcc accatattgc cgtcttttgg caatgtgagg gcccggaaac ctggccctgt
     3181 cttcttgacg agcattccta ggggtctttc ccctctcgcc aaaggaatgc aaggtctgtt
     3241 gaatgtcgtg aaggaagcag ttcctctgga agtttcttga agacaaacaa cgtctgtagc
     3301 gaccctttgc aggcagcgga accccccacc tggcgacagg tgcctctgcg gccaaaagcc
     3361 acgtgtataa gatacacctg caaaggcggc acaaccccag tgccacgttg tgagttggat
     3421 agttgtggaa agagtcaaat ggctctcctc aagcgtattc aacaaggggc tgaaggatgc
     3481 ccagaaggta ccccattgta tgggatctga tctggggcct cggtgcacat gctttacatg
     3541 tgtttagtcg aggttaaaaa aacgtctagg ccccccgaac cacggggacg tggttttcct
     3601 ttgaaaaaca cgatgataat atggccacaa ccgtcgacat gaccgagtac aagcccacgg
     3661 tgcgcctcgc cacccgcgac gacgtccccc gggccgtacg caccctcgcc gccgcgttcg
     3721 ccgactaccc cgccacgcgc cacaccgtcg acccggaccg ccacatcgag cgggtcaccg
     3781 agctgcaaga actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg
     3841 acgacggcgc cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt
     3901 tcgccgagat cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac
     3961 agatggaagg cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg
     4021 tcggcgtctc gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag
     4081 tggaggcggc cgagcgcgcc ggggtgcccg ccttcctgga gacctccgcg ccccgcaacc
     4141 tccccttcta cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac
     4201 cgcgcacctg gtgcatgacc cgcaagcccg gtgcctgagt cgaccgcccg ccccacgacc
     4261 cgcagcgccc gaccgaaagg agcgcacgac cccatggctc gacgataaaa taaaagattt
     4321 tatttagtct ccagaaaaag gggggaatga aagaccccac ctgtaggttt ggcaagctag
     4381 cttaagtaac gccattttgc aaggcatgga aaaatacata actgagaata gagaagttca
     4441 gatcaaggtc aggaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     4501 gcagttcctg ccccggctca gggccaagaa cagatggaac agctgaatat gggccaaaca
     4561 ggatatctgt ggtaagcagt tcctgccccg gctcagggcc aagaacagat ggtccccaga
     4621 tgcggtccag ccctcagcag tttctagaga accatcagat gtttccaggg tgccccaagg
     4681 acctgaaatg accctgtgcc ttatttgaac taaccaatca gttcgcttct cgcttctgtt
     4741 cgcgcgcttc tgctccccga gctcaataaa agagcccaca acccctcact cggggcgcca
     4801 gtcctccgat tgactgagtc gcccgggtac ccgtgtatcc aataaaccct cttgcagttg
     4861 catccgactt gtggtctcgc tgttccttgg gagggtctcc tctgagtgat tgactacccg
     4921 tcagcggggg tctttcacat gcagcatgta tcaaaattaa tttggttttt tttcttaagt
     4981 atttacatta aatggccata gttgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     5041 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     5101 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     5161 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     5221 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     5281 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     5341 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     5401 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     5461 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     5521 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     5581 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     5641 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     5701 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     5761 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     5821 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     5881 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttgcgg
     5941 ccggccgcaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     6001 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     6061 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     6121 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     6181 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     6241 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     6301 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     6361 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     6421 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     6481 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     6541 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     6601 cgtcaacacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     6661 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6721 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6781 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6841 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6901 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6961 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     7021 aaaataggcg tatcacgagg ccctttcgtc ttcactatag gagcttggcc cattgcatac
     7081 gttgtatcca tatcataata tgtacattta tattggctca tgtccaacat taccgccatg
     7141 ttgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     7201 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     7261 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     7321 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     7381 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     7441 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     7501 attagtcatc gctattacca tgctagagaa ccatcagatg tttccagggt gccccaagga
     7561 cctgaaatga ccctgtgcct tatttgaact aaccaatcag ttcgcttctc gcttctgttc
     7621 gcgcgcttct gctccccgag ctcaataaaa gagcccacaa cccctcactc ggcgcgccag
     7681 tcctccgatt gactgagtcg cccgggtacc cgtgtatcca ataaaccctc ttgcag
//
