LOCUS       Exported                7575 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  MAC-tagged gene expression.
ACCESSION   .
VERSION     .
KEYWORDS    MAC_C_LAMP1
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7575)
  AUTHORS   Liu X, Salokas K, Tamene F, Jiu Y, Weldatsadik RG, Ohman T, 
            Varjosalo M
  TITLE     An AP-MS- and BioID-compatible MAC-tag enables comprehensive mapping
            of protein interactions and subcellular localizations.
  JOURNAL   Nat Commun. 2018 Mar 22;9(1):1188. doi: 10.1038/s41467-018-03523-2.
  PUBMED    29568061
REFERENCE   2  (bases 1 to 7575)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7575
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        7..386
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        387..590
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     541..561
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    592..610
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    613..631
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     641..665
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    808..832
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             832..2082
                     /codon_start=1
                     /product="human lysosome-associated membrane glycoprotein 
                     1"
                     /label=LAMP1
                     /translation="MAAPGSARRPLLLLLLLLLLGLMHCASAAMFMVKNGNGTACIMAN
                     FSAAFSVNYDTKSGPKNMTFDLPSDATVVLNRSSCGKENTSDPSLVIAFGRGHTLTLNF
                     TRNATRYSVQLMSFVYNLSDTHLFPNASSKEIKTVESITDIRADIDKKYRCVSGTQVHM
                     NNVTVTLHDATIQAYLSNSSFSRGETRCEQDRPSPTTAPPAPPSPSPSPVPKSPSVDKY
                     NVSGTNGTCLLASMGLQLNLTYERKDNTTVTRLLNINPNKTSASGSCGAHLVTLELHSE
                     GTTVLLFQFGMNASSSRFFLQGIQLNTILPDARDPAFKAANGSLRALQATVGNSYKCNA
                     EEHVRVTKAFSVNIFKVWVQAFKVEGGQFGSVEECLLDENSMLIPIAVGGALAGLVLIV
                     LIAYLVGRKRSHAGYQTI"
     protein_bind    complement(2084..2108)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2155..2178
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2215..2238
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             2269..2295
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2296..3255
                     /codon_start=1
                     /product="promiscuous R118G mutant of the E. coli biotin 
                     protein ligase (Cronan, 2005)"
                     /label=BirA(R118G)
                     /note="also known as BirA* or BioID"
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGGRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     primer_bind     complement(3299..3316)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3305..3529
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3662..3681)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     protein_bind    3813..3860
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             3868..4890
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    5020..5141
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5057..5076)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5111..5130
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5190..5206)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5190..5206)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5203..5225)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5214..5230
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5238..5268)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5283..5304
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5421..5438)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5592..6180)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5672..5691)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6351..7211)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6974..6993
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7212..7316)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7391..7410)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 cgcgttgaca ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc
       61 atagcccata tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac
      121 cgcccaacga cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa
      181 tagggacttt ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag
      241 tacatcaagt gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc
      301 ccgcctggca ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct
      361 acgtattagt catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg
      421 gatagcggtt tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt
      481 tgttttggca ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga
      541 cgcaaatggg cggtaggcgt gtacggtggg aggtctatat aagcagagct ctccctatca
      601 gtgatagaga tctccctatc agtgatagag atcgtcgacg agctcgttta gtgaaccgtc
      661 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      721 ccagcctccg gactctagcg tttaaactta agcttggtac cgagctcgga tccactagtc
      781 cagtgtggtg gaattctgca gatatcaaca agtttgtaca aaaaagttgg catggcggcc
      841 cccggcagcg cccggcgacc cctgctgctg ctactgctgt tgctgctgct cggcctcatg
      901 cattgtgcgt cagcagcaat gtttatggtg aaaaatggca acgggaccgc gtgcataatg
      961 gccaacttct ctgctgcctt ctcagtgaac tacgacacca agagtggccc taagaacatg
     1021 acctttgacc tgccatcaga tgccacagtg gtgctcaacc gcagctcctg tggaaaagag
     1081 aacacttctg accccagtct cgtgattgct tttggaagag gacatacact cactctcaat
     1141 ttcacgagaa atgcaacacg ttacagcgtc cagctcatga gttttgttta taacttgtca
     1201 gacacacacc ttttccccaa tgcgagctcc aaagaaatca agactgtgga atctataact
     1261 gacatcaggg cagatataga taaaaaatac agatgtgtta gtggcaccca ggtccacatg
     1321 aacaacgtga ccgtaacgct ccatgatgcc accatccagg cgtacctttc caacagcagc
     1381 ttcagcaggg gagagacacg ctgtgaacaa gacaggcctt ccccaaccac agcgccccct
     1441 gcgccaccca gcccctcgcc ctcacccgtg cccaagagcc cctctgtgga caagtacaac
     1501 gtgagcggca ccaacgggac ctgcctgctg gccagcatgg ggctgcagct gaacctcacc
     1561 tatgagagga aggacaacac gacggtgaca aggcttctca acatcaaccc caacaagacc
     1621 tcggccagcg ggagctgcgg cgcccacctg gtgactctgg agctgcacag cgagggcacc
     1681 accgtcctgc tcttccagtt cgggatgaat gcaagttcta gccggttttt cctacaagga
     1741 atccagttga atacaattct tcctgacgcc agagaccctg cctttaaagc tgccaacggc
     1801 tccctgcgag cgctgcaggc cacagtcggc aattcctaca agtgcaacgc ggaggagcac
     1861 gtccgtgtca cgaaggcgtt ttcagtcaat atattcaaag tgtgggtcca ggctttcaag
     1921 gtggaaggtg gccagtttgg ctctgtggag gagtgtctgc tggacgagaa cagcatgctg
     1981 atccccatcg ctgtgggtgg tgccctggcg gggctggtcc tcatcgtcct catcgcctac
     2041 ctcgtcggca ggaagaggag tcacgcaggc taccagacta tctacccaac tttcttgtac
     2101 aaagtggttg atatcaaggc cgccgacatc accagcctgt acaagaaagt ggggtggagc
     2161 catccccagt tcgagaaggg cggaggatct ggcggcggaa gcggcggagg aagctggtcc
     2221 caccctcagt ttgagaaagg caccgagctg ggcagcacca tggccagcta cccctacgac
     2281 gtgcccgact acgccaagga caacaccgtg cccctgaagc tgatcgccct gctggccaac
     2341 ggcgagttcc actctggcga gcagctgggc gagacactgg gcatgagcag agccgccatc
     2401 aacaagcaca tccagaccct gcgggactgg ggcgtggacg tgttcacagt gcccggcaag
     2461 ggctacagcc tgcccgagcc catccagctg ctgaacgcca agcagatcct gggccagctg
     2521 gatggcggat ctgtggccgt gctgcccgtg atcgacagca ccaaccagta tctgctcgac
     2581 cggatcggcg agctgaagtc cggcgacgcc tgtatcgccg agtaccagca ggccggcaga
     2641 ggcggcaggg gcagaaagtg gttcagcccc ttcggcgcca acctgtacct gagcatgttc
     2701 tggagactgg aacagggccc agccgccgct attggcctga gcctggtcat cggcatcgtg
     2761 atggccgagg tgctgcggaa gctgggcgcc gacaaagtgc gcgtgaagtg gcccaacgac
     2821 ctgtatctgc aggaccggaa gctggccggc atcctggtgg aactgaccgg caaaacaggc
     2881 gacgccgccc agatcgtgat cggcgctggc atcaacatgg ccatgcggcg ggtggaagag
     2941 agcgtggtca accagggctg gatcaccctg caggaagccg gcatcaacct ggaccggaac
     3001 accctggccg ccatgctgat cagagagctg agagccgccc tggaactgtt cgagcaggaa
     3061 ggcctggccc cctacctgag cagatgggag aagctggaca acttcatcaa ccggcccgtg
     3121 aaactgatca tcggcgacaa agagatcttc ggcatcagcc ggggcatcga caagcagggg
     3181 gccctgctcc tggaacagga cggcatcatc aagccctgga tgggcggcga gatcagcctg
     3241 cggagcgccg agaagtgatg actcgagtct agagggcccg tttaaacccg ctgatcagcc
     3301 tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg
     3361 accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat
     3421 tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag
     3481 gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg
     3541 gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc
     3601 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     3661 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     3721 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     3781 aaacttgatt agggtgatgg ttcacgtacc tagaagttcc tattccgaag ttcctattct
     3841 ctagaaagta taggaacttc cttggccaaa aagcctgaac tcaccgcgac gtctgtcgag
     3901 aagtttctga tcgaaaagtt cgacagcgtc tccgacctga tgcagctctc ggagggcgaa
     3961 gaatctcgtg ctttcagctt cgatgtagga gggcgtggat atgtcctgcg ggtaaatagc
     4021 tgcgccgatg gtttctacaa agatcgttat gtttatcggc actttgcatc ggccgcgctc
     4081 ccgattccgg aagtgcttga cattggggaa ttcagcgaga gcctgaccta ttgcatctcc
     4141 cgccgtgcac agggtgtcac gttgcaagac ctgcctgaaa ccgaactgcc cgctgttctg
     4201 cagccggtcg cggaggccat ggatgcgatc gctgcggccg atcttagcca gacgagcggg
     4261 ttcggcccat tcggaccgca aggaatcggt caatacacta catggcgtga tttcatatgc
     4321 gcgattgctg atccccatgt gtatcactgg caaactgtga tggacgacac cgtcagtgcg
     4381 tccgtcgcgc aggctctcga tgagctgatg ctttgggccg aggactgccc cgaagtccgg
     4441 cacctcgtgc acgcggattt cggctccaac aatgtcctga cggacaatgg ccgcataaca
     4501 gcggtcattg actggagcga ggcgatgttc ggggattccc aatacgaggt cgccaacatc
     4561 ttcttctgga ggccgtggtt ggcttgtatg gagcagcaga cgcgctactt cgagcggagg
     4621 catccggagc ttgcaggatc gccgcggctc cgggcgtata tgctccgcat tggtcttgac
     4681 caactctatc agagcttggt tgacggcaat ttcgatgatg cagcttgggc gcagggtcga
     4741 tgcgacgcaa tcgtccgatc cggagccggg actgtcgggc gtacacaaat cgcccgcaga
     4801 agcgcggccg tctggaccga tggctgtgta gaagtactcg ccgatagtgg aaaccgacgc
     4861 cccagcactc gtccgagggc aaaggaatag cacgtactac gagatttcga ttccaccgcc
     4921 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4981 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     5041 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     5101 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     5161 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     5221 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     5281 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     5341 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     5401 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     5461 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     5521 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     5581 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     5641 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     5701 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     5761 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     5821 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     5881 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     5941 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6001 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     6061 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     6121 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     6181 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     6241 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     6301 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     6361 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     6421 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     6481 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     6541 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     6601 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     6661 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     6721 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     6781 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     6841 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     6901 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     6961 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7021 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7081 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     7141 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     7201 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     7261 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     7321 tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt
     7381 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
     7441 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
     7501 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
     7561 tacgggccag atata
//
