LOCUS       sequence-120709-        8247 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Ecadherin reporter only (no hook) (RUSH system).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8247)
  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 8247)
  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: SBP-mCherry-Ecadherin_puromycin
FEATURES             Location/Qualifiers
     source          1..8247
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     sig_peptide     945..1004
                     /codon_start=1
                     /product="human interleukin-2 signal sequence"
                     /label=IL-2 signal sequence
                     /translation="MYRMQLLSCIALSLALVTNS"
     CDS             1005..1115
                     /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="DEKTTGWRGGHVVEGLAGELEQLRARLEHHPQGQREP"
     CDS             1125..1832
                     /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(1270..1288)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1538..1557
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     intron          4126..4355
                     /label=chimeric intron
                     /note="chimera between introns from adenovirus and 
                     immunoglobulin heavy chain genes"
     primer_bind     4338..4357
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     misc_feature    join(4428..5002,5003..5005,5006..5014)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4595..4612)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4822..4841
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5034..5633
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(5034..5053)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5530..5550
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    5867..5988
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5904..5923)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6090..6106)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6090..6106)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6103..6125)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6114..6130
                     /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(6138..6144,6145..6162,6163..6168))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6183..6204
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6321..6338)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6492..7080)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6572..6591)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7251..8042,8043..8111))
                     /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     7874..7893
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8112..8216)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gagctcggat cgatatctgc ggcctagcta gcgggaggcg cgccatgtac aggatgcaac
      961 tcctgtcttg cattgcacta agtcttgcac ttgtcacgaa ttccgacgag aagaccactg
     1021 gttggcgagg tggacacgtt gttgaaggac tggctgggga acttgaacaa cttcgtgcac
     1081 gactggagca tcacccacaa ggtcaacgtg aaccacctgc aggtatggtg agcaagggcg
     1141 aggaggataa catggccatc atcaaggagt tcatgcgctt caaggtgcac atggagggct
     1201 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     1261 cccagaccgc caagctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     1321 tgtcccctca gttcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     1381 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     1441 acggcggcgt ggtgaccgtg acccaggact cctccctaca ggacggcgag ttcatctaca
     1501 aggtgaagct gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     1561 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     1621 tcaagcagag gctgaagctg aaggacggcg gccactacga cgctgaggtc aagaccacct
     1681 acaaggccaa gaagcccgtg cagctgcccg gcgcctacaa cgtcaacatc aagttggaca
     1741 tcacctccca caacgaggac tacaccatcg tggaacagta cgaacgcgcc gagggccgcc
     1801 actccaccgg cggcatggac gagctgtaca agggccggcc agactgggtc atccctccca
     1861 tcagctgccc cgaaaatgaa aagggtgaat tcccaaagaa cctggttcag atcaaatcca
     1921 acagggacaa agaaacaaag gttttctaca gcatcaccgg ccaaggagct gacaaacccc
     1981 ccgttggcgt tttcatcatt gagagggaga caggctggct gaaagtgaca cagcctctgg
     2041 atagagaagc cattgccaag tacatcctct attctcatgc cgtgtcatca aatggggaag
     2101 cggtggagga tcccatggag atagtgatca cagtgacaga tcagaatgac aacaggccag
     2161 agtttaccca ggaggtgttt gagggatccg ttgcagaagg cgctgttcca ggaacctccg
     2221 tgatgaaggt ctcagccacc gatgcagacg atgacgtcaa cacctacaac gctgccatcg
     2281 cctacaccat cgtcagccag gatcctgagc tgcctcacaa aaacatgttc actgtcaata
     2341 gggacaccgg ggtcatcagt gtgctcacct ctgggctgga ccgagagagt taccctacat
     2401 acactctggt ggttcaggct gctgaccttc aaggcgaagg cttgagcaca acagccaagg
     2461 ctgtgatcac tgtcaaggat attaatgaca acgctcctgt cttcaacccg agcacgtatc
     2521 agggtcaagt gcctgagaat gaggtcaatg cccggatcgc cacactcaaa gtgaccgatg
     2581 atgatgcccc caacactccg gcgtggaaag ctgtgtacac cgtagtcaac gatcctgacc
     2641 agcagttcgt tgttgtcaca gaccccacga ccaatgatgg cattttgaaa acagccaagg
     2701 gcttggattt tgaggccaag cagcaataca tccttcatgt gagagtggag aacgaggaac
     2761 cctttgaggg gtctcttgtc ccttccacag ccactgtcac tgtggacgtg gtagacgtga
     2821 atgaagcccc catctttatg cctgcggaga ggagagtcga agtgcccgaa gactttggtg
     2881 tgggtcagga aatcacatct tataccgctc gagagccgga cacgttcatg gatcagaaga
     2941 tcacgtatcg gatttggagg gacactgcca actggctgga gattaaccca gagactggtg
     3001 ccattttcac gcgcgctgag atggacagag aagacgctga gcatgtgaag aacagcacat
     3061 atgtagctct catcatcgcc acagatgatg gttcacccat tgccactggc acgggcactc
     3121 ttctcctggt cctgttagac gtcaatgaca acgctcccat cccagaacct cgaaacatgc
     3181 agttctgcca gaggaaccca cagcctcata tcatcaccat cttggatcca gaccttcccc
     3241 ccaacacgtc cccctttact gctgagctaa cccatggggc cagcgtcaac tggaccattg
     3301 agtataatga cgcagctcaa gaatctctca ttttgcaacc aagaaaggac ttagagattg
     3361 gcgaatacaa aatccatctc aagctcgcgg ataaccagaa caaagaccag gtgaccacgt
     3421 tggacgtcca tgtgtgtgac tgtgaaggga cggtcaacaa ctgcatgaag gcgggaatcg
     3481 tggcagcagg attgcaagtt cctgccatcc tcggaatcct tggagggatc ctcgccctgc
     3541 tgattctgat cctgctgctc ctactgtttc tacggaggag aacggtggtc aaagagcccc
     3601 tgctgccacc agatgatgat acccgggaca atgtgtatta ctatgatgaa gaaggaggtg
     3661 gagaagaaga ccaggacttt gatttgagcc agctgcacag gggcctggat gcccgaccgg
     3721 aagtgactcg aaatgatgtg gctcccaccc tcatgagcgt gccccagtat cgtccccgtc
     3781 ctgccaatcc tgatgaaatt ggaaacttca tcgatgaaaa cctgaaggca gccgacagcg
     3841 accccacggc acccccttac gactctctgt tggtgttcga ttacgagggc agtggttctg
     3901 aagccgctag cctgagctca ctgaactcct ctgagtcgga tcaggaccag gactacgatt
     3961 atctgaacga gtggggcaac cgattcaaga agctggcgga catgtacggc ggtggcgaag
     4021 acgactaggg cctcgagggc caattaatta aggatccgcg gccgcataga taactgatcc
     4081 agtgtgctgg aattaattcg ctgtctgcga gggccagctg ttggggtgag tactccctct
     4141 caaaagcggg catgacttct gcgctaagat tgtcagtttc caaaaacgag gaggatttga
     4201 tattcacctg gcccgcggtg atgcctttga gggtggccgc gtccatctgg tcagaaaaga
     4261 caatcttttt gttgtcaagc ttgaggtgtg gcaggcttga gatctggcca tacacttgag
     4321 tgacaatgac atccactttg cctttctctc cacaggtgtc cactcccagg tccaactgca
     4381 ggtcgagcat gcatctaggg cggccaattc cgcccctctc cccccccccc ctctccctcc
     4441 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     4501 atgtgatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     4561 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     4621 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     4681 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     4741 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     4801 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     4861 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     4921 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     4981 ctttgaaaaa cacgatgata agcttgccac aacccacaag gagacgacct tccatgaccg
     5041 agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt cccccgggcc gtacgcaccc
     5101 tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgacccg gaccgccaca
     5161 tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca
     5221 aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg
     5281 aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc
     5341 tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt
     5401 ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg
     5461 tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct
     5521 ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg
     5581 aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc
     5641 cccacgaccc gcagcgcccg accgaaagga gcgcacgacc ccatggctcc gaccgaagcc
     5701 acccggggcg gccccgccga ccccgcaccc gcccccgagg cccaccgact ctagataact
     5761 gatcataatc agccatacca catttgtaga ggttttactt gctttaaaaa acctcccaca
     5821 cctccccctg aacctgaaac ataaaatgaa tgcaattgtt gttgttaact tgtttattgc
     5881 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     5941 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttaac gcgtcgagtg
     6001 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     6061 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     6121 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6181 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6241 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6301 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6361 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6421 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6481 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6541 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6601 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6661 ccttcgggaa gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag
     6721 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6781 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     6841 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     6901 aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg
     6961 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7021 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7081 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7141 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7201 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7261 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7321 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7381 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7441 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7501 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7561 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7621 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7681 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7741 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7801 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     7861 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     7921 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     7981 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8041 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8101 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     8161 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     8221 tttccccgaa aagtgccacc tgacgtc
//
