LOCUS       Exported                7386 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synchronize trafficking of ManII from the ER (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    Ii-Str_ManII-SBP-tagBFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7386)
  AUTHORS   Boncompain G, Divoux S, Gareil N, de Forges H, Lescure A, Latreche 
            L, Mercanti V, Jollivet F, Raposo G, Perez F
  TITLE     Synchronization of secretory protein traffic in populations of 
            cells.
  JOURNAL   Nat Methods. 2012 Mar 11;9(5):493-8. doi: 10.1038/nmeth.1928.
  PUBMED    22406856
REFERENCE   2  (bases 1 to 7386)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7386
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             1401..1427
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          1946..2175
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2158..2177
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2233..2806
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2400..2417)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2627..2646
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3208..3321
                     /codon_start=1
                     /product="streptavidin-binding peptide"
                     /label=SBP
                     /note="selected from a peptide library; binds streptavidin 
                     with nanomolar affinity (Keefe et al., 2001)"
                     /translation="MDEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     polyA_signal    4174..4295
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4211..4230)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4265..4284
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4302..4757)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4439..4460)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4651..4670
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4784..4888
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4890..5247
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4913..4933)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5098..5233
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5160..5179
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             5282..6076
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(5336..5355)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5946..5965
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6264..6283)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6308..6355
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      6684..7272
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7173..7192
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcgcta
      601 ccggtcgtac gtctccggat tcgaattaat tcggatccgt gatgcacagg aggagagcca
      661 gggcctgtcg ggaagatcaa aagccagtca tcgatgatca gcgcgacctt atctccaaca
      721 atgagcaact gcccatgctg ggccggcgcc ctggggcccc ggagagcaag tgcagccgcg
      781 gagccctgta cacaggcttt tccatcctgg tgactctgct cctcgctggc caggccacca
      841 ccgcctactt cctgtaccag cagcagggcc ggctggacaa actgacagtc acctcccaga
      901 acctgcagct cgagatggac cctagcaaag actcaaaagc tcaggtgtcc gctgccgagg
      961 ctggcattac tggaacatgg tacaatcagc tcgggagcac ctttattgtg actgctggag
     1021 ccgatggagc cctcaccgga acatacgaat ctgctgtggg aaacgccgaa tcacggtacg
     1081 tcctcactgg ccgatacgat agtgcccctg ccaccgacgg atctgggact gccctgggat
     1141 ggactgtcgc ttggaaaaac aactaccgga atgctcattc tgccacaaca tggagtggac
     1201 agtacgtggg aggcgctgag gctagaatca atacacagtg gctgctcaca tctggcacaa
     1261 ccgaggcaaa tgcttggaaa tccaccctgg tgggacatga cacattcacc aaagtgaaac
     1321 cctccgccgc ttcaatcgat gccgccaaaa aagccggagt caacaacggc aatcctctgg
     1381 atgccgtcca gcaggtcgac tatccgtacg acgtaccaga ctacgcactc gacctgcagc
     1441 tggagaacct gcgcatgaag cttcccaagc ctcccaagcc tgtgagcaag atgcgcatgg
     1501 ccaccccgct gctgatgcat gcgctgccca tgggagccct gccccagggg cccatgcaga
     1561 atgccaccaa gtatggcaac atgacagagg accatgtgat gcacctgctc cagaatgctg
     1621 accccctgaa ggtgtacccg ccactgaagg ggagcttccc ggagaacctg agacacctta
     1681 agaacaccat ggagaccata gactggaagg tctttgagag ctggatgcac cattggctcc
     1741 tgtttgaaat gagcaggcac tccttggagc aaaagcccac tgacgctcca ccgaaagagt
     1801 cactggaact ggaggacccg tcttctgggc tgggtgtgac caagcaggat ctgggcccag
     1861 tccccatgtg aggatccgcg gccgcataga taactgatcc agtgtgctgg aattaattcg
     1921 ctgtctgcga gggccagctg ttggggtgag tactccctct caaaagcggg catgacttct
     1981 gcgctaagat tgtcagtttc caaaaacgag gaggatttga tattcacctg gcccgcggtg
     2041 atgcctttga gggtggccgc gtccatctgg tcagaaaaga caatcttttt gttgtcaagc
     2101 ttgaggtgtg gcaggcttga gatctggcca tacacttgag tgacaatgac atccactttg
     2161 cctttctctc cacaggtgtc cactcccagg tccaactgca ggtcgagcat gcatctaggg
     2221 cggccaattc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga
     2281 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa
     2341 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc
     2401 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc
     2461 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg
     2521 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca
     2581 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag
     2641 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct
     2701 ggggcctcgg tgcacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc
     2761 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataagctt gccacaaccc
     2821 gggaggcgcg ccatgaagtt aagtcgccag ttcaccgtgt ttggcagcgc gatcttctgc
     2881 gtcgtaatct tctcactcta cctgatgctg gacaggggtc acttggacta ccctcggggc
     2941 ccgcgccagg agggctcctt tccgcagggc cagctttcaa tattgcaaga aaagattgac
     3001 catttggagc gtttgctcgc tgagaacaac gagattatct caaatatcag agactcagtc
     3061 atcaacctga gcgagtctgt ggaggacggc ccgcgggggt caccaggcaa cgccagccaa
     3121 ggctccatcc acctccactc gccacagttg gccctgcagg ctgaccccag agactgtttg
     3181 gatcccaccg gtcgccaccg gaattccatg gacgagaaga ccactggttg gcgaggtgga
     3241 cacgttgttg aaggactggc tggggaactt gaacaacttc gtgcacgact ggagcatcac
     3301 ccacaaggtc aacgtgaacc acctgcaggt tctgaactca tcaaggaaaa catgcatatg
     3361 aaactgtaca tggagggaac cgtggataat caccacttca agtgtactag tgagggcgag
     3421 ggcaaacctt acgagggaac acagaccatg aggatcaaag tcgtcgaggg cggacctctg
     3481 cctttcgctt tcgacattct ggccacatcc tttctgtacg gctccaaaac cttcatcaac
     3541 cacacacagg gcatccccga tttcttcaaa cagtccttcc ccgagggctt cacatgggaa
     3601 cgagtgacaa cctacgagga cggaggagtg ctcaccgcta ctcaggatac tagcctccag
     3661 gatggatgtc tcatctacaa cgtgaaaatc cgaggcgtga acttcacctc aaatggcccc
     3721 gtgatgcaga aaaaaactct cggctgggag gcctttactg aaaccctgta ccctgccgat
     3781 ggcggcctgg aggggcgaaa tgatatggct ctgaaactcg tgggaggatc tcacctcatt
     3841 gccaacatca aaaccaccta ccggagcaaa aaacctgcca aaaatctgaa aatgcctggc
     3901 gtctactacg tcgattaccg gctggaacga atcaaagagg ccaacaacga aacttacgtg
     3961 gaacagcacg aagtcgccgt ggctcgatac tgtgacctgc ctagcaaact gggacacaaa
     4021 ggccggcctt aaggcctcga gggccaatta attaactcta gataactgat cataatcagc
     4081 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     4141 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     4201 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     4261 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     4321 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     4381 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     4441 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     4501 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     4561 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     4621 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     4681 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     4741 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4801 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4861 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     4921 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     4981 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     5041 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     5101 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     5161 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     5221 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     5281 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     5341 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5401 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5461 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5521 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5581 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5641 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5701 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5761 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     5821 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5881 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5941 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     6001 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     6061 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     6121 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     6181 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     6241 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     6301 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     6361 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     6421 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     6481 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     6541 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     6601 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     6661 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     6721 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     6781 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     6841 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     6901 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6961 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     7021 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     7081 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     7141 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     7201 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     7261 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     7321 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     7381 atgcat
//
