LOCUS       Exported                6404 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Cytoplasmic ER hook only (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    Str-Ii_neomycin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  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 6404)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6404
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(48..67)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        239..618
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        619..822
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     773..793
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     867..886
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        867..885
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1459..1485
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2220..2449
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2432..2451
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    2507..3080
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2674..2691)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2901..2920
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3107..3901
                     /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(3161..3180)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3771..3790
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4028..4149
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4065..4084)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4251..4267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4251..4267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4264..4286)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4275..4291
                     /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(4299..4329)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4344..4365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4482..4499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4653..5241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4733..4752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(5412..6272)
                     /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     6035..6054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6273..6377)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cgtcgacgga tcgggagatc tcccgatccc ctatggtcga ctctcagtac aatctgctct
       61 gatgccgcat agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag
      121 tgcgcgagca aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat
      181 ctgcttaggg ttaggcgttt tgcgctgctt cgcgatgtac gggccagata tacgcgttga
      241 cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt tcatagccca
      301 tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
      361 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
      421 ttccattgac gtcaatgggt ggactattta cggtaaactg cccacttggc agtacatcaa
      481 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
      541 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
      601 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
      661 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
      721 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
      781 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctctctggct aactagagaa
      841 cccactgctt actggcttat cgaaattaat acgactcact atagggagac ccaagcttgg
      901 taccgagctc ggatcgatat catgcaccgg aggagatcac gctcttgtag ggaggaccag
      961 aaacctgtca ccggtgaccc tagcaaagac tcaaaagctc aggtgtccgc tgccgaggct
     1021 ggcattactg gaacatggta caatcagctc gggagcacct ttattgtgac tgctggagcc
     1081 gatggagccc tcaccggaac atacgaatct gctgtgggaa acgccgaatc acggtacgtc
     1141 ctcactggcc gatacgatag tgcccctgcc accgacggat ctgggactgc cctgggatgg
     1201 actgtcgctt ggaaaaacaa ctaccggaat gctcattctg ccacaacatg gagtggacag
     1261 tacgtgggag gcgctgaggc tagaatcaat acacagtggc tgctcacatc tggcacaacc
     1321 gaggcaaatg cttggaaatc caccctggtg ggacatgaca cattcaccaa agtgaaaccc
     1381 tccgccgctt caatcgatgc cgccaaaaaa gccggagtca acaacggcaa tcctctggat
     1441 gccgtccagc aggtcgacta tccgtacgac gtaccagact acgcagtcgg accgatggac
     1501 gatcagaggg acctcattag caacaacgaa cagctgccta tgctgggacg gcgacctgga
     1561 gcccctgaat ccaaatgctc taggggagca ctgtacactg gcttctccat tctcgtgaca
     1621 ctgctgctgg ccgggcaggc tactactgct tacttcctgt accagcagca ggggcggctg
     1681 gacaaactca ctgtgacatc tcagaacctc cagctggaaa atctgaggat gaaactgccc
     1741 aaacccccta aacccgtgtc caaaatgagg atggccacac ctctgctcat gcaggcactg
     1801 ccaatgggag ccctgcccca ggggcccatg cagaatgcca ccaagtatgg caacatgaca
     1861 gaggaccatg tgatgcacct gctccagaat gctgaccccc tgaaggtgta cccgccactg
     1921 aaggggagct tcccggagaa cctgagacac cttaagaaca ccatggagac catagactgg
     1981 aaggtctttg agagctggat gcaccattgg ctcctgtttg aaatgagcag gcactccttg
     2041 gagcaaaagc ccactgacgc tccaccgaaa gagtcactgg aactggagga cccgtcttct
     2101 gggctgggtg tgaccaagca ggatctgggc ccagtcccca tgtgaggatc cgcggccgca
     2161 tagataactg atccagtgtg ctggaattaa ttcgctgtct gcgagggcca gctgttgggg
     2221 tgagtactcc ctctcaaaag cgggcatgac ttctgcgcta agattgtcag tttccaaaaa
     2281 cgaggaggat ttgatattca cctggcccgc ggtgatgcct ttgagggtgg ccgcgtccat
     2341 ctggtcagaa aagacaatct ttttgttgtc aagcttgagg tgtggcaggc ttgagatctg
     2401 gccatacact tgagtgacaa tgacatccac tttgcctttc tctccacagg tgtccactcc
     2461 caggtccaac tgcaggtcga gcatgcatct agggcggcca attccgcccc tctccctccc
     2521 ccccccctaa cgttactggc cgaagccgct tggaataagg ccggtgtgcg tttgtctata
     2581 tgttattttc caccatattg ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg
     2641 tcttcttgac gagcattcct aggggtcttt cccctctcgc caaaggaatg caaggtctgt
     2701 tgaatgtcgt gaaggaagca gttcctctgg aagcttcttg aagacaaaca acgtctgtag
     2761 cgaccctttg caggcagcgg aaccccccac ctggcgacag gtgcctctgc ggccaaaagc
     2821 cacgtgtata agatacacct gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga
     2881 tagttgtgga aagagtcaaa tggctctcct caagcgtatt caacaagggg ctgaaggatg
     2941 cccagaaggt accccattgt atgggatctg atctggggcc tcggtgcaca tgctttacat
     3001 gtgtttagtc gaggttaaaa aaacgtctag gccccccgaa ccacggggac gtggttttcc
     3061 tttgaaaaac acgatgataa gcttgccaca acccgggagg cgcgccatga ttgaacaaga
     3121 tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc
     3181 acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc
     3241 ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc
     3301 gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac
     3361 tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc
     3421 tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac
     3481 gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg
     3541 tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct
     3601 cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt
     3661 cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg
     3721 attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac
     3781 ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg
     3841 tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg
     3901 attaattaac tctagataac tgatcataat cagccatacc acatttgtag aggttttact
     3961 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     4021 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4081 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4141 tgtatcttaa cgcgtcgagt gcattctagt tgtggtttgt ccaaactcat caatgtatct
     4201 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     4261 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4321 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4381 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     4441 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     4501 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4561 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4621 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     4681 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     4741 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     4801 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt
     4861 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     4921 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     4981 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5041 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     5101 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5161 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5221 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5281 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5341 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5401 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5461 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5521 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5581 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     5641 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     5701 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     5761 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     5821 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     5881 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     5941 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6001 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6061 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6121 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6181 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6241 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6301 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6361 caaatagggg ttccgcgcac atttccccga aaagtgccac ctga
//
