LOCUS       sequence-242449-        6340 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  express Siah1 RM.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6340)
  TITLE     Ronai lab expression plasmids
REFERENCE   2  (bases 1 to 6340)
  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"
COMMENT     Sequence Label: pcDNA3-Flag-Siah1 Ring Mutant
FEATURES             Location/Qualifiers
     source          1..6340
                     /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"
     regulatory      905..914
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             914..937
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(1893..1911)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1894..1911)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1931..1948)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1937..2161
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2207..2635
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2294..2313)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2504..2525
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2644..2664)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2649..2978
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2829..2964
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2891..2910
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3045..3839
                     /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(3099..3118)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3709..3728
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4013..4134
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4050..4069)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4104..4123
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4183..4199)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4183..4199)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4196..4218)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4207..4223
                     /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(4231..4237,4238..4255,4256..4261))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4276..4297
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4414..4431)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4585..5173)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4665..4684)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5344..6135,6136..6204))
                     /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     5967..5986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6205..6309)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ggagacccta gaactagtgg
      901 atccgccacc atggactaca aggacgacga tgacaaagat cccccgatct ctcgaggatc
      961 cgaattcccg cccacagaga tgagccgcca gactgctaca gcattaccca ctggcacctc
     1021 aaagtgtcca ccatcccaga gggtacctgc cttgaccggc acaactgcat ccaacaatga
     1081 cttggcgagt ctttttgagt gtcctgtctg ctttgactat gtgttgccac ctattcttca
     1141 gtgtcagagt ggcgctcttg ttgctagcaa ctgtcgcccc aaacttacat gttgtcccac
     1201 ttgccggggc ccattgggat ccattcgcaa cttggctatg gagaaagtgg ccaactcagt
     1261 actcttccct tgtaaatatg cctcttctgg atgtgaaata actctgccac acaccgaaaa
     1321 ggcagagcac gaggagctct gtgagttcag gccttactcc tgcccctgcc ctggtgcttc
     1381 ctgtaagtgg caaggctcct tggatgccgt catgccccac ctgatgcatc agcacaagtc
     1441 cattaccacc ctgcaaggag aagatatagt tttccttgct acagacatta accttcctgg
     1501 tgctgttgac tgggtgatga tgcagtcttg ttttggcttt catttcatgt tagtcttgga
     1561 gaaacaagaa aaatatgatg gtcatcagca gttctttgca attgtacaac tgataggaac
     1621 acgcaagcaa gctgaaaatt ttgcatatcg acttgagcta aatggtcata ggcggcgatt
     1681 gacttgggaa gcgactcctc ggtctattca tgagggaatt gcaacagcca ttatgaatag
     1741 tgactgccta gtgtttgaca ccagcattgc acagcttttt gcagaaaatg gcaatttagg
     1801 catcaatgta actatttcca tgtgttgaaa cggcaatcaa ctcgagtcag ccgaattcgc
     1861 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     1921 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1981 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     2041 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2101 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2161 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2221 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2281 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2341 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2401 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2461 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2521 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2581 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2641 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2701 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2761 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccrcccctaa
     2821 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2881 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2941 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3001 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3061 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3121 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3181 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     3241 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3301 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     3361 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     3421 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     3481 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     3541 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     3601 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     3661 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     3721 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     3781 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     3841 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     3901 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     3961 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ccaacttgtt
     4021 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     4081 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt
     4141 ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc tgtttcctgt
     4201 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     4261 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     4321 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcggggag
     4381 aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt
     4441 cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga
     4501 atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg
     4561 taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa
     4621 aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     4681 tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct
     4741 gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct
     4801 cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc
     4861 cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt
     4921 atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc
     4981 tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat
     5041 ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa
     5101 acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa
     5161 aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga
     5221 aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct
     5281 tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga
     5341 cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     5401 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     5461 ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat
     5521 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     5581 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     5641 caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     5701 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     5761 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     5821 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     5881 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     5941 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     6001 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     6061 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     6121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     6181 gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca
     6241 gggttattgt ctcatgagcg aatacatatt tgaatgtatt tagaaaaata aacaaatagg
     6301 ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
