LOCUS       sequence-110464-        6920 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synchronize trafficking of ManII from the ER.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6920)
  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 6920)
  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"; doi: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2012-03-11- 11"; volume: "9"; 
            issue: "5"; pages: "493-8"
COMMENT     Sequence Label: Ii-Str_ManII-SBP-mCherry
FEATURES             Location/Qualifiers
     source          1..6920
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             1753..1779
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     intron          2298..2527
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     2510..2529
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(2585..3159,3160..3162,3163..3171)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2752..2769)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     2979..2998
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             3560..3673
                     /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"
     CDS             3683..4390
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3828..3846)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     4096..4115
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    4544..4665
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4581..4600)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(4767..4783)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4767..4783)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4780..4802)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4791..4807
                     /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(join(4815..4821,4822..4839,4840..4845))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4860..4881
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4998..5015)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5169..5757)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5249..5268)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5928..6719,6720..6788))
                     /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     6551..6570
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6789..6893)
                     /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 ctgcggccta gctagcgctt aaggcctgtt aaccggtcgt
      961 acgtctccgg attcgaatta attcggatcc gtgatgcaca ggaggagagc cagggcctgt
     1021 cgggaagatc aaaagccagt catcgatgat cagcgcgacc ttatctccaa caatgagcaa
     1081 ctgcccatgc tgggccggcg ccctggggcc ccggagagca agtgcagccg cggagccctg
     1141 tacacaggct tttccatcct ggtgactctg ctcctcgctg gccaggccac caccgcctac
     1201 ttcctgtacc agcagcaggg ccggctggac aaactgacag tcacctccca gaacctgcag
     1261 ctcgagatgg accctagcaa agactcaaaa gctcaggtgt ccgctgccga ggctggcatt
     1321 actggaacat ggtacaatca gctcgggagc acctttattg tgactgctgg agccgatgga
     1381 gccctcaccg gaacatacga atctgctgtg ggaaacgccg aatcacggta cgtcctcact
     1441 ggccgatacg atagtgcccc tgccaccgac ggatctggga ctgccctggg atggactgtc
     1501 gcttggaaaa acaactaccg gaatgctcat tctgccacaa catggagtgg acagtacgtg
     1561 ggaggcgctg aggctagaat caatacacag tggctgctca catctggcac aaccgaggca
     1621 aatgcttgga aatccaccct ggtgggacat gacacattca ccaaagtgaa accctccgcc
     1681 gcttcaatcg atgccgccaa aaaagccgga gtcaacaacg gcaatcctct ggatgccgtc
     1741 cagcaggtcg actatccgta cgacgtacca gactacgcac tcgacctgca gctggagaac
     1801 ctgcgcatga agcttcccaa gcctcccaag cctgtgagca agatgcgcat ggccaccccg
     1861 ctgctgatgc atgcgctgcc catgggagcc ctgccccagg ggcccatgca gaatgccacc
     1921 aagtatggca acatgacaga ggaccatgtg atgcacctgc tccagaatgc tgaccccctg
     1981 aaggtgtacc cgccactgaa ggggagcttc ccggagaacc tgagacacct taagaacacc
     2041 atggagacca tagactggaa ggtctttgag agctggatgc accattggct cctgtttgaa
     2101 atgagcaggc actccttgga gcaaaagccc actgacgctc caccgaaaga gtcactggaa
     2161 ctggaggacc cgtcttctgg gctgggtgtg accaagcagg atctgggccc agtccccatg
     2221 tgaggatccg cggccgcata gataactgat ccagtgtgct ggaattaatt cgctgtctgc
     2281 gagggccagc tgttggggtg agtactccct ctcaaaagcg ggcatgactt ctgcgctaag
     2341 attgtcagtt tccaaaaacg aggaggattt gatattcacc tggcccgcgg tgatgccttt
     2401 gagggtggcc gcgtccatct ggtcagaaaa gacaatcttt ttgttgtcaa gcttgaggtg
     2461 tggcaggctt gagatctggc catacacttg agtgacaatg acatccactt tgcctttctc
     2521 tccacaggtg tccactccca ggtccaactg caggtcgagc atgcatctag ggcggccaat
     2581 tccgcccctc tccctccccc ccccctaacg ttactggccg aagccgcttg gaataaggcc
     2641 ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc gtcttttggc aatgtgaggg
     2701 cccggaaacc tggccctgtc ttcttgacga gcattcctag gggtctttcc cctctcgcca
     2761 aaggaatgca aggtctgttg aatgtcgtga aggaagcagt tcctctggaa gcttcttgaa
     2821 gacaaacaac gtctgtagcg accctttgca ggcagcggaa ccccccacct ggcgacaggt
     2881 gcctctgcgg ccaaaagcca cgtgtataag atacacctgc aaaggcggca caaccccagt
     2941 gccacgttgt gagttggata gttgtggaaa gagtcaaatg gctctcctca agcgtattca
     3001 acaaggggct gaaggatgcc cagaaggtac cccattgtat gggatctgat ctggggcctc
     3061 ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa acgtctaggc cccccgaacc
     3121 acggggacgt ggttttcctt tgaaaaacac gatgataagc ttgccacaac ccgggaggcg
     3181 cgccatgaag ttaagtcgcc agttcaccgt gtttggcagc gcgatcttct gcgtcgtaat
     3241 cttctcactc tacctgatgc tggacagggg tcacttggac taccctcggg gcccgcgcca
     3301 ggagggctcc tttccgcagg gccagctttc aatattgcaa gaaaagattg accatttgga
     3361 gcgtttgctc gctgagaaca acgagattat ctcaaatatc agagactcag tcatcaacct
     3421 gagcgagtct gtggaggacg gcccgcgggg gtcaccaggc aacgccagcc aaggctccat
     3481 ccacctccac tcgccacagt tggccctgca ggctgacccc agagactgtt tggatcccac
     3541 cggtcgccac cggaattcca tggacgagaa gaccactggt tggcgaggtg gacacgttgt
     3601 tgaaggactg gctggggaac ttgaacaact tcgtgcacga ctggagcatc acccacaagg
     3661 tcaacgtgaa ccacctgcag gtatggtgag caagggcgag gaggataaca tggccatcat
     3721 caaggagttc atgcgcttca aggtgcacat ggagggctcc gtgaacggcc acgagttcga
     3781 gatcgagggc gagggcgagg gccgccccta cgagggcacc cagaccgcca agctgaaggt
     3841 gaccaagggt ggccccctgc ccttcgcctg ggacatcctg tcccctcagt tcatgtacgg
     3901 ctccaaggcc tacgtgaagc accccgccga catccccgac tacttgaagc tgtccttccc
     3961 cgagggcttc aagtgggagc gcgtgatgaa cttcgaggac ggcggcgtgg tgaccgtgac
     4021 ccaggactcc tccctacagg acggcgagtt catctacaag gtgaagctgc gcggcaccaa
     4081 cttcccctcc gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga
     4141 gcggatgtac cccgaggacg gcgccctgaa gggcgagatc aagcagaggc tgaagctgaa
     4201 ggacggcggc cactacgacg ctgaggtcaa gaccacctac aaggccaaga agcccgtgca
     4261 gctgcccggc gcctacaacg tcaacatcaa gttggacatc acctcccaca acgaggacta
     4321 caccatcgtg gaacagtacg aacgcgccga gggccgccac tccaccggcg gcatggacga
     4381 gctgtacaag ggccggcctt aaggcctcga gggccaatta attaactcta gataactgat
     4441 cataatcagc cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct
     4501 ccccctgaac ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc
     4561 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     4621 actgcattct agttgtggtt tgtccaaact catcaatgta tcttaacgcg tcgagtgcat
     4681 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4741 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4801 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4861 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4921 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4981 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5041 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5101 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5161 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5221 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5281 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5341 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5401 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5461 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5521 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5581 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5641 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5701 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5761 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5821 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5881 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5941 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6001 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6061 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6121 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6181 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6241 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6301 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6361 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6421 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6481 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6541 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6601 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6661 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6721 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6781 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6841 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6901 ccccgaaaag tgccacctga
//
