LOCUS       sequence-227758-        5751 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human SUMO3 with HA tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5751)
  AUTHORS   Kamitani T, Nguyen HP, Kito K, Fukuda-Kamitani T, Yeh ET
  TITLE     Covalent modification of PML by the sentrin family of ubiquitin-like
            proteins.
  JOURNAL   J Biol Chem. 1998 Feb 6. 273(6):3117-20.
  PUBMED    9452416
REFERENCE   2  (bases 1 to 5751)
  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: "J Biol 
            Chem. 1998 Feb 6. 273(6):3117-20."
COMMENT     Sequence Label: pcDNA3/HA-SUMO3 (Sentrin 2)
FEATURES             Location/Qualifiers
     source          1..5751
                     /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"
     CDS             898..924
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             934..1209
                     /codon_start=1
                     /product="human small ubiquitin-related modifier 3"
                     /label=SUMO3
                     /translation="MSEEKPKEGVKTENDHINLKVAGQDGSVVQFKIKRHTPLSKLMKA
                     YCERQGLSMRQIRFRFDGQPINETDTPAQLEMEDEDTIDVFQQQTGG"
     promoter        complement(1304..1322)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1305..1322)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1342..1359)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1348..1572
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1618..2046
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1705..1724)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1915..1936
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2055..2075)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2060..2389
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2240..2375
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2302..2321
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2456..3250
                     /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(2510..2529)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3120..3139
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3424..3545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3461..3480)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3515..3534
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3594..3610)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3594..3610)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3607..3629)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3618..3634
                     /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(3642..3648,3649..3666,3667..3672))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3687..3708
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3825..3842)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3996..4584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4076..4095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4755..5546,5547..5615))
                     /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     5378..5397
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5616..5720)
                     /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 gcttatgtac
      901 ccatacgatg ttccggatta cgctagcgga tccatgtccg aggagaagcc caaggagggt
      961 gtgaagacag agaatgacca catcaacctg aaggtggccg ggcaggacgg ctccgtggtg
     1021 cagttcaaga tcaagaggca cacgccgctg agcaagctga tgaaggccta ctgcgagagg
     1081 cagggcttgt caatgaggca gatcagattc aggttcgacg ggcagccaat caatgaaact
     1141 gacactccag cacagctgga gatggaggac gaggacacca tcgacgtgtt ccagcagcag
     1201 acgggaggtg tgccggagag cagcctggca gggcacagtt tctgaattct gcagatatcc
     1261 atcacactgg cggccgctcg agcatgcatc tagagggccc tattctatag tgtcacctaa
     1321 atgctagagc tcgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     1381 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     1441 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     1501 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     1561 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     1621 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     1681 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     1741 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     1801 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     1861 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     1921 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     1981 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     2041 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     2101 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     2161 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     2221 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     2281 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     2341 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     2401 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     2461 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     2521 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     2581 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     2641 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     2701 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     2761 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     2821 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     2881 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     2941 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     3001 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     3061 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     3121 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     3181 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     3241 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     3301 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     3361 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     3421 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     3481 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     3541 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     3601 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     3661 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     3721 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     3781 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     3841 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     3901 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     3961 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4021 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4081 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4141 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4201 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4261 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4321 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4381 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4441 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4501 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4561 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     4621 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     4681 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     4741 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     4801 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     4861 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     4921 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     4981 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5041 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5101 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5161 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5221 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5281 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5341 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5401 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5461 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5521 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     5581 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     5641 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     5701 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
