LOCUS       sequence-271745-        7249 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of SH3BP1R12A in mammalian cells; resistant to siRNA 
            targeting GAUGACAGCCACCCACUUC.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7249)
  AUTHORS   Elbediwy A, Zihni C, Terry SJ, Clark P, Matter K, Balda MS
  TITLE     Epithelial junction formation requires confinement of Cdc42 activity
            by a novel SH3BP1 complex.
  JOURNAL   J Cell Biol. 2012 Aug 20;198(4):677-93. doi: 10.1083/jcb.201202094. 
            Epub 2012 Aug 13.
  PUBMED    22891260
REFERENCE   2  (bases 1 to 7249)
  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: 
            "10.1083/jcb.201202094"; journalName: "J Cell Biol"; date: 
            "2012-08-20- 20"; volume: "198"; issue: "4"; pages: "677-93"
COMMENT     Sequence Label: pcDNA-TO-HA-SH3BP1 R312A (RNAi-res)
FEATURES             Location/Qualifiers
     source          1..7249
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     protein_bind    586..604
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    607..625
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     635..659
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             2938..2967
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2983..3000
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3023..3040)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3029..3253
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3299..3727
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3386..3405)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3596..3617
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3736..3756)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3741..4070
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3921..4056
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3983..4002
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4118..4165
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4184..4558
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4688..4809
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4725..4744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4779..4798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4858..4874)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4858..4874)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4871..4893)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4882..4898
                     /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(4906..4912,4913..4930,4931..4936))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4951..4972
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5089..5106)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5260..5848)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5340..5359)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6019..6810,6811..6879))
                     /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     6642..6661
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6880..6984)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7059..7078)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctccct atcagtgata
      601 gagatctccc tatcagtgat agagatcgtc gacgagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccggactct agcgtttaaa cttaagcttg gtaccgagct cggatccatg caggacctgc
      781 caggcaacga caacagcacc gccggcctcg agatgatgaa gaggcagctg caccgcatgc
      841 ggcagctggc ccagacgggc agcttgggac gcaccccgga gaccgctgag ttcctgggtg
      901 aggacctgct gcaggtagaa cagcggctgg agccggccaa gcgggcagcc cacaacatcc
      961 acaagcggct gcaggcctgt ctgcagggcc agagcggggc agacatggac aagcgggtga
     1021 agaagcttcc cctcatggct ctgtccacca cgatggctga gagcttcaag gagctggacc
     1081 ctgattccag catggggaag gccttggaga tgagctgtgc catccagaat cagctggccc
     1141 gcatcctggc cgagtttgag atgaccctgg agagggacgt cctgcagcca ctcagcaggc
     1201 tgagtgagga ggagctgcca gccatcctca aacacaagaa aagcctccag aagctcgtgt
     1261 ccgactggaa cacactcaag agcaggctca gtcaggcaac caagaattca ggcagcagtc
     1321 aaggcctagg aggcagcccg ggtagtcaca gccatacgac catggccaac aaggtggaga
     1381 cgctgaagga ggaggaggag gagctgaaga ggaaagtgga gcaatgcagg gacgagtact
     1441 tggctgacct gtaccacttt gttaccaagg aggactccta tgccaactac ttcattcgtc
     1501 tcctggagat tcaggccgat taccatcgca ggtcactgag ctcgctggac acagccctgg
     1561 ctgagctgag ggagaaccac ggccaagcag accactcccc ttcaatgaca gcaacacact
     1621 tccccagggt gtatggggtg tcgctggcaa cccacctgca agagctgggc cgggagattg
     1681 ccctgcccat cgaggcctgc gtcatgatgc tgctttctga gggcatgaag gaagagggtc
     1741 tcttcgctct ggctgctggg gcctcggtgc tgaagcgtct caagcagaca atggcctcgg
     1801 acccccacag cctggaggag ttctgctccg acccgcacgc tgtggcaggt gccctcaagt
     1861 cctatctgcg ggagctgcca gagcctctga tgaccttcga cctctatgat gactggatga
     1921 gggcagccag cctgaaggag ccaggggccc ggctgcaggc cctccaagag gtgtgcagcc
     1981 gcctaccccc cgagaacctc agcaacctca ggtacctgat gaagttcctg gcacggctgg
     2041 ccgaggagca ggaggtgaac aagatgacac ccagcaacat cgccatagtc ctgggaccca
     2101 acttgctgtg gccacctgag aaagaagggg accaggccca gctggatgca gcctccgtgt
     2161 cttccatcca ggtggtgggc gtcgtcgagg cgctgatcca gagcgcagac accctcttcc
     2221 ctggagacat caacttcaac gtgtcaggcc tcttctcagc tgttaccctc caggacacag
     2281 tcagtgacag gctggcctct gaggaacttc cgtccactgc cgtgcccacc ccagccacca
     2341 ccccggctcc ggctccggct ccagctccag ctccggcccc agccttggct tcagcagcta
     2401 ccaaggaaag gacagagtct gaggtgcctc ccagaccagc ctcccccaag gtcaccagga
     2461 gtcccccgga gacagctgcc ccagtggagg acatggctcg gaggaccaag cgcccggcgc
     2521 cagcccggcc caccatgccg cccccccagg tctccggctc ccgctcctcc cctccagccc
     2581 cgcccttgcc ccctggctct ggcagccctg ggacccccca agccctgccc cgacgtctgg
     2641 ttggcagcag cctccgagcc cccacagtgc cacccccgtt accccccaca ccccctcagc
     2701 ctgcccggcg ccaaagccgg cgttcaccag cctcccccag cccggcctcc ccaggtccag
     2761 cctcccccag cccagtctct ttgagtaacc ctgcacaggt ggacctgggg gctgccacag
     2821 cagagggagg agcccctgag gctatcagtg gggtccccac tcccccagct atccccccgc
     2881 agccccgccc caggagcctt gcctcagaga ccaactgatc tagagggccc gcggttcgaa
     2941 caaaaactca tctcagaaga ggatctgaat atgcataccg gtcatcatca ccatcaccat
     3001 tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     3061 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     3121 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     3181 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     3241 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     3301 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     3361 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     3421 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3481 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3541 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3601 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3661 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3721 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3781 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3841 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3901 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3961 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     4021 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     4081 gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat catcggcata
     4141 gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca agttgaccag
     4201 tgccgttccg gtgctcaccg cgcgcgacgt cgccggagcg gtcgagttct ggaccgaccg
     4261 gctcgggttc tcccgggact tcgtggagga cgacttcgcc ggtgtggtcc gggacgacgt
     4321 gaccctgttc atcagcgcgg tccaggacca ggtggtgccg gacaacaccc tggcctgggt
     4381 gtgggtgcgc ggcctggacg agctgtacgc cgagtggtcg gaggtcgtgt ccacgaactt
     4441 ccgggacgcc tccgggccgg ccatgaccga gatcggcgag cagccgtggg ggcgggagtt
     4501 cgccctgcgc gacccggccg gcaactgcgt gcacttcgtg gccgaggagc aggactgaca
     4561 cgtgctacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4621 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4681 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4741 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4801 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4861 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4921 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4981 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     5041 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     5101 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     5161 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     5221 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     5281 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     5341 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     5401 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5461 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5521 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5581 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5641 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5701 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5761 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5821 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5881 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5941 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     6001 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     6061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     6121 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     6181 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     6241 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     6301 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     6361 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6421 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6481 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6541 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6601 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6661 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6721 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6781 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6841 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6901 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6961 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     7021 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     7081 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     7141 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     7201 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
