LOCUS       sequence-343425-        7972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses Numb in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7972)
  AUTHORS   Yu CH, Rafiq NB, Cao F, Zhou Y, Krishnasamy A, Biswas KH, Ravasio A,
            Chen Z, Wang YH, Kawauchi K, Jones GE, Sheetz MP
  TITLE     Integrin-beta3 clusters recruit clathrin-mediated endocytic 
            machinery in the absence of traction force.
  JOURNAL   Nat Commun. 2015 Oct 28;6:8672. doi: 10.1038/ncomms9672.
  PUBMED    26507506
REFERENCE   2  (bases 1 to 7972)
  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"; journalName: "Nat 
            Commun."; date: "2015-10-28"; volume: "6:8672. doi"; pages: " 
            10.1038/ncomms9672"
COMMENT     Sequence Label: Numb-mCherry
FEATURES             Location/Qualifiers
     source          1..7972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        116..495
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        496..707
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     658..678
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     704..728
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          868..1000
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     983..1002
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1045..1063
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1080..1106
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2892..3602
                     /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(3037..3055)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3305..3324
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    3654..3775
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3665..3684)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3719..3738
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     rep_origin      3961..4416
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4048..4067)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4258..4279
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        4533..4890
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4533..4724
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4556..4576)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4741..4876
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4941..5735
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4995..5014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5605..5624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5799..5847
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(5888..5907)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6007..6029
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6067..6085)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6153..6257
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6258..6326,6327..7118)
                     /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     complement(6476..6495)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7289..7877
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7778..7797
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca tacgttgtat
       61 ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc atgttggcat
      121 tgattattga ctagttatta atagtaatca attacggggt cattagttca tagcccatat
      181 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      241 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      301 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      361 tatcatatgc caagtccgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      421 tatgcccagt acatgacctt acgggacttt cctacttggc agtacatcta cgtattagtc
      481 atcgctatta ccatggtgat gcggttttgg cagtacacca atgggcgtgg atagcggttt
      541 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      601 caaaatcaac gggactttcc aaaatgtcgt aacaactgcg atcgcccgcc ccgttgacgc
      661 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctcgt ttagtgaacc
      721 gtcagatcac tagaagcttt attgcggtag tttatcacag ttaaattgct aacgcagtca
      781 gtgcttctga cacaacagtc tcgaacttaa gctgcagtga ctctcttaag gtagccttgc
      841 agaagttggt cgtgaggcac tgggcaggta agtatcaagg ttacaagaca ggtttaagga
      901 gaccaataga aactgggctt gtcgagacag agaagactct tgcgtttctg ataggcacct
      961 attggtctta ctgacatcca ctttgccttt ctctccacag gtgtccactc ccagttcaat
     1021 tacagctctt aaggctagag tacttaatac gactcactat aggctaggcc cccaacatgt
     1081 acccatacga cgtcccagac tacgctctcg aggaattcac gcgtttcagg agaaagaaag
     1141 acgtttatgt cccagaggcc agccgtccac atcagtggca gacagatgaa gaaggagtcc
     1201 gcactggaaa gtgtagcttc ccagttaagt acctcggcca cgtagaagtt gatgagtcaa
     1261 gaggaatgca catctgtgaa gatgccgtaa agagattgaa agctacggga aagaaagcag
     1321 tgaaggccgt tctgtgggtg tcagcggatg ggctcagagt tgtggacgag aaaactaagg
     1381 acctcatagt tgaccagaca atagaaaaag tttccttctg tgccccagat aggaactttg
     1441 acagagcctt ttcttacata tgtcgcgatg gcaccactcg gcgatggatc tgtcattgtt
     1501 tcatggctgt caaagacacg ggggaaagac tgagccacgc cgtgggctgt gcttttgcag
     1561 cctgtttaga gcgtaaacag aagcgggaga aggagtgtgg cgtcactgct acttttgatg
     1621 ccagtagaac cactttcaca agagaaggat cattccgtgt cacaactgcc actgagcaag
     1681 ccgaaagaga ggagatcatg aaacagttgc aagatgccaa gaaagctgag acagacaaga
     1741 cagctgttgg tccatcagtg gctcctggca acactgctcc atccccatcc tctcccacct
     1801 ctcccactcc ggatggcact gcatcttcag agatgaacaa tccccatgct atcccacgcc
     1861 ggcatgcacc aattgaacag cttgctcgtc aaggctcttt ccggggattt cctgctctta
     1921 gccagaagat gtcacccttt aaacgccagc tgtccctccg catcaatgag ttgccttcca
     1981 ctatgcagag gaagactgat ttcccaataa aaaacacagt gcccgaggtg gaaggagagg
     2041 ccgagagcat cagctccttg tgttcccaga tcaccagtgc cttcagcacg ccctctgagg
     2101 accccttctc ctccgcccca atgaccaaac cagtgacatt ggtggcacca cagtctcctg
     2161 tgttacaagg gactgagtgg ggtcagtctt ctggtgctgc ctctccaggt ctcttccagg
     2221 ctggtcacag acgcactccc tctgaagctg accgctggtt agaagaagtg tcaaagagtg
     2281 tgcgggccca gcagcctcag gtctcagctg cccctctgca gccagttctc cagccgcctc
     2341 cgcccgccgc cattgcccct ccagcacctc ctttccaagg acatgccttc ctcacgtccc
     2401 agcctgtgcc cgtgggtgtg gtcccacccc tacaaccagc ctttgtccct acccagtcct
     2461 accctgtggc caacgggatg ccctacccag cctctaatgt gcctgtagtg ggcatcaccc
     2521 catcccagat ggtagccaat gtgtttggca ctgcaggcca ccctcagacc actcatccac
     2581 atcagtcgcc aagcctggcc aagcagcaga cattccctca atatgagaca agtagtgcta
     2641 ccaccagtcc cttctttaag cctcctgctc agcacctcaa tggttctgca gctttcaatg
     2701 gtgtagacaa tggtgggcta gcctcaggaa acaggcatgc agaagtccct ccaggcacct
     2761 gcccagtgga tcctttcgaa gctcagtggg ccgcactaga aagcaagtcc aagcagcgta
     2821 caaatccttc tcctaccaac cctttctcca gtgacttaca gaaaacattt gaaatagaac
     2881 ttcgacccgg gatggtgagc aagggcgagg aggataacat ggccatcatc aaggagttca
     2941 tgcgcttcaa ggtgcacatg gagggctccg tgaacggcca cgagttcgag atcgagggcg
     3001 agggcgaggg ccgcccctac gagggcaccc agaccgccaa gctgaaggtg accaagggtg
     3061 gccccctgcc cttcgcctgg gacatcctgt cccctcagtt catgtacggc tccaaggcct
     3121 acgtgaagca ccccgccgac atccccgact acttgaagct gtccttcccc gagggcttca
     3181 agtgggagcg cgtgatgaac ttcgaggacg gcggcgtggt gaccgtgacc caggactcct
     3241 ccctgcagga cggcgagttc atctacaagg tgaagctgcg cggcaccaac ttcccctccg
     3301 acggccccgt aatgcagaag aagaccatgg gctgggaggc ctcctccgag cggatgtacc
     3361 ccgaggacgg cgccctgaag ggcgagatca agcagaggct gaagctgaag gacggcggcc
     3421 actacgacgc tgaggtcaag accacctaca aggccaagaa gcccgtgcag ctgcccggcg
     3481 cctacaacgt caacatcaag ttggacatca cctcccacaa cgaggactac accatcgtgg
     3541 aacagtacga acgcgccgag ggccgccact ccaccggcgg catggacgag ctgtacaagt
     3601 aacccgggcg gccgcttccc tttagtgagg gttaatgctt cgagcagaca tgataagata
     3661 cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct ttatttgtga
     3721 aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac aagttaacaa
     3781 caacaattgc attcatttta tgtttcaggt tcagggggag atgtgggagg ttttttaaag
     3841 caagtaaaac ctctacaaat gtggtaaaat ccgataagga tcgatccggg ctggcgtaat
     3901 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     3961 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     4021 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     4081 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     4141 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     4201 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     4261 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     4321 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     4381 acgcgaattt taacaaaata ttaacgctta caatttcctg atgcggtatt ttctccttac
     4441 gcatctgtgc ggtatttcac accgcatacg cggatctgcg cagcaccatg gcctgaaata
     4501 acctctgaaa gaggaacttg gttaggtacc ttctgaggcg gaaagaacca gctgtggaat
     4561 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     4621 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga
     4681 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc
     4741 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt
     4801 tttatttatg cagaggccga ggccgcctcg gcctctgagc tattccagaa gtagtgagga
     4861 ggcttttttg gaggcctagg cttttgcaaa aagcttgatt cttctgacac aacagtctcg
     4921 aacttaaggc tagagccacc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4981 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     5041 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     5101 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     5161 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5221 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5281 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5341 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5401 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5461 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5521 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5581 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5641 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5701 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5761 cgaccaagcg acgcccaacc tgccatcacg atggccgcaa taaaatatct ttattttcat
     5821 tacatctgtg tgttggtttt ttgtgtgaat cgatagcgat aaggatccgc gtatggtgca
     5881 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     5941 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     6001 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     6061 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     6121 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6181 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6241 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6301 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6361 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6421 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6481 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     6541 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6601 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6661 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6721 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6781 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6841 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     6901 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     6961 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7021 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7081 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7141 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7201 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7261 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     7321 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     7381 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     7441 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     7501 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     7561 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     7621 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     7681 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     7741 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     7801 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     7861 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     7921 ggccttttgc tcacatggct cgacagatct tcaatattgg ccattagcca ta
//
