LOCUS       Exported                7895 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses FAM134B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    pMRX-INU-FLAG-FAM134B
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7895)
  AUTHORS   Chino H, Hatta T, Natsume T, Mizushima N
  TITLE     Intrinsically Disordered Protein TEX264 Mediates ER-phagy.
  JOURNAL   Mol Cell. 2019 Apr 11. pii: S1097-2765(19)30257-6. doi: 
            10.1016/j.molcel.2019.03.033.
  PUBMED    31006538
REFERENCE   2  (bases 1 to 7895)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7895
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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             3512..3577
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    3627..4179
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3771..3788)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3998..4017
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             4197..4991
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4251..4270)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4861..4880
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     LTR             5221..5690
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(5820..5837)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5991..6579)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6071..6090)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6743..7603)
                     /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     7366..7385
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7604..7708)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7776..7794
                     /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 cgagcccggc gcctccggag cacgccgagg agggatgccc
     2041 ggctcctgcc gccgaggagc aggcgccgcc gtcgccgcca ccgccccagg catcccccgc
     2101 agagcggcag cagcaggagg aggaagcgca ggaagctggg gcggcggagg gcgcggggtt
     2161 gcaggtggag gaggccgcgg gccgggcggc ggccgcggta acctggctgc tcggggagcc
     2221 ggtgctgtgg ctgggctgcc gcgccgacga gctgctgagc tggaagaggc cgctgcggag
     2281 cctgctcggc ttcgtcgctg ccaacctgct gttctggttc cttgcattga ctccatggag
     2341 agtatatcac ctgatttccg tcatgatact tgggcgtgtt attatgcaaa taataaagga
     2401 tatggttttg tctagaacaa gaggtgcaca gttgtggaga agcctcagtg aaagctggga
     2461 agttatcaat tccaaaccag atgaaagacc caggctcagc cactgtattg cagaatcatg
     2521 gatgaatttc agcatatttc ttcaagaaat gtctcttttt aaacagcaga gccctggcaa
     2581 gttttgtctc ctggtctgta gtgtgtgcac attttttacg atcttgggaa gttacattcc
     2641 tggggttata ctcagctatc tactgttact gtgtgcattt ttgtgtccat tgtttaaatg
     2701 taatgatatt ggacaaaaaa tttacagcaa aattaagtca gttctgctga aactggattt
     2761 tggaattgga gaatatatta atcagaagaa acgtgagaga tctgaagcag acaaagaaaa
     2821 aagtcacaaa gatgacagtg aattagactt ttcagctctt tgtcctaaga ttagcctcac
     2881 ggttgctgcc aaagagttat ctgtgtctga cacagacgtc tcagaggtat cctggactga
     2941 taatgggacc ttcaaccttt cagaaggata cactccacag acagacactt ctgatgatct
     3001 tgaccgaccc agtgaggaag ttttctctag agatctttca gattttccat ctctagaaaa
     3061 tggcatggga acaaatgatg aagatgaatt aagccttggt ttgcccactg agctcaagag
     3121 aaagaaggaa cagttggaca gtggtcacag accaagcaaa gagacgcaat cagcagctgg
     3181 tctcaccctt cctctgaaca gtgaccaaac ctttcacctg atgagcaacc tggctgggga
     3241 tgttatcaca gctgcagtga ctgcagctat caaagaccag ttagagggtg tgcagcaagc
     3301 actttctcag gctgccccca tcccagaaga ggacacagac actgaagaag gtgatgactt
     3361 tgaactactt gaccagtcag agctggatca aattgagagt gaattgggac ttacacaaga
     3421 ccaggaagca gaagcacagc aaaataagaa gtcttcaggt ttcctttcaa atctgctggg
     3481 aggccatggg aattcgggct ctggttcagg cgactacaaa gaccatgacg gtgattataa
     3541 agatcatgac atcgactaca aggatgacga tgacaagtaa tgatttctcg agagcggccg
     3601 ctacgtaaat tccgcccccc cccctaacgt tactggccga agccgcttgg aataaggccg
     3661 gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc
     3721 ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa
     3781 aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag tttcttgaag
     3841 acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg
     3901 cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg
     3961 ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa
     4021 caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg
     4081 gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca
     4141 cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc gtcgacatga
     4201 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4261 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4321 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     4381 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4441 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4501 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4561 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4621 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4681 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     4741 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     4801 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     4861 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     4921 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     4981 agttcttctg agtcgaccgc ccgccccacg acccgcagcg cccgaccgaa aggagcgcac
     5041 gaccccatgg ctcgacgata aaataaaaga ttttatttag tctccagaaa aaggggggaa
     5101 tgaaagaccc cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat
     5161 ggaaaaatac ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag
     5221 ctgaatatgg gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa
     5281 gaacagatgg aacagctgaa tatgggccaa acaggatatc tgtggtaagc agttcctgcc
     5341 ccggctcagg gccaagaaca gatggtcccc agatgcggtc cagccctcag cagtttctag
     5401 agaaccatca gatgtttcca gggtgcccca aggacctgaa atgaccctgt gccttatttg
     5461 aactaaccaa tcagttcgct tctcgcttct gttcgcgcgc ttctgctccc cgagctcaat
     5521 aaaagagccc acaacccctc actcggggcg ccagtcctcc gattgactga gtcgcccggg
     5581 tacccgtgta tccaataaac cctcttgcag ttgcatccga cttgtggtct cgctgttcct
     5641 tgggagggtc tcctctgagt gattgactac ccgtcagcgg gggtctttca catgcagcat
     5701 gtatcaaaat taatttggtt ttttttctta agtatttaca ttaaatggcc atagttgcat
     5761 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     5821 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     5881 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     5941 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     6001 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     6061 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     6121 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     6181 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     6241 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     6301 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     6361 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     6421 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     6481 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     6541 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     6601 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     6661 tcaaaaagga tcttcaccta gatccttttg cggccggccg caaatcaatc taaagtatat
     6721 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     6781 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     6841 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     6901 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     6961 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     7021 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     7081 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     7141 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     7201 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     7261 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     7321 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaac acgggataat accgcgccac
     7381 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     7441 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     7501 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     7561 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     7621 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     7681 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgtct
     7741 aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg aggccctttc
     7801 gtcttcacta taggagcttg gcccattgca tacgttgtat ccatatcata atatgtacat
     7861 ttatattggc tcatgtccaa cattaccgcc atgtt
//
