LOCUS       Exported                5823 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression of SUMO2 with a strep and an HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3.1-StrepHA-SUMO2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5823)
  AUTHORS   Bursomanno S, McGouran JF, Kessler BM, Hickson ID, Liu Y
  TITLE     Regulation of SUMO2 target proteins by the proteasome in human cells
            exposed to replication stress.
  JOURNAL   J Proteome Res. 2015 Apr 3;14(4):1687-99. doi: 10.1021/pr500997p. 
            Epub 2015 Mar 19.
  PUBMED    25748227
REFERENCE   2  (bases 1 to 5823)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5823
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     CDS             932..955
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             992..1015
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     CDS             1043..1069
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1085..1363
                     /codon_start=1
                     /product="human small ubiquitin-related modifier 2"
                     /label=SUMO2
                     /translation="MADEKPKEGVKTENNDHINLKVAGQDGSVVQFKIKRHTPLSKLMK
                     AYCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG"
     primer_bind     complement(1417..1434)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1423..1647
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1693..2121
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1780..1799)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1990..2011
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2130..2150)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2135..2464
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2315..2450
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2377..2396
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2531..3325
                     /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(2585..2604)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3195..3214
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3499..3620
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3536..3555)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3590..3609
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3669..3685)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3669..3685)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3682..3704)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3693..3709
                     /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(3717..3747)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3762..3783
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3900..3917)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4071..4656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4151..4170)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4827..5687)
                     /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     5450..5469
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5688..5792)
                     /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 ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccatggctag ctggagccac ccgcagttcg agaaaggtgg
      961 aggttccgga ggtggatcgg gaggtggatc gtggagccac ccgcagttcg aaaaaggcgc
     1021 cgagaccgcg gtcccgaatt cttaccctta tgacgtcccc gattacgccg ctgcagcgga
     1081 tatcatggcc gacgaaaagc ccaaggaagg agtcaagact gagaacaacg atcatattaa
     1141 tttgaaggtg gcggggcagg atggttctgt ggtgcagttt aagattaaga ggcatacacc
     1201 acttagtaaa ctaatgaaag cctattgtga acgacaggga ttgtcaatga ggcagatcag
     1261 attccgattt gacgggcaac caatcaatga aacagacaca cctgcacagt tggaaatgga
     1321 ggatgaagat acaattgatg tgttccaaca gcagacggga ggtgtctact gagcggccgc
     1381 tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct tctagttgcc
     1441 agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt gccactccca
     1501 ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg tgtcattcta
     1561 ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac aatagcaggc
     1621 atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc tggggctcta
     1681 gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     1741 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     1801 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     1861 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     1921 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     1981 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     2041 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     2101 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     2161 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     2221 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2281 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     2341 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     2401 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     2461 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga caggatgagg
     2521 atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg cttgggtgga
     2581 gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt
     2641 ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt ccggtgccct
     2701 gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg gcgttccttg
     2761 cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat tgggcgaagt
     2821 gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat ccatcatggc
     2881 tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg accaccaagc
     2941 gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg atcaggatga
     3001 tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgcg
     3061 catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat
     3121 ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg
     3181 ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc
     3241 tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta
     3301 tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac cgaccaagcg
     3361 acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     3421 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     3481 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     3541 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     3601 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     3661 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     3721 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     3781 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat
     3841 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     3901 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3961 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4021 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4081 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4141 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4201 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4261 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4321 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4381 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4441 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4501 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4561 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt ttttgtttgc
     4621 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4681 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4741 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4801 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4861 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4921 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4981 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     5041 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     5101 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5161 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5221 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5281 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5341 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5401 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5461 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5521 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5581 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5641 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5701 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5761 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5821 gtc
//
