LOCUS       sequence-72029-e        5127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of N-terminally tagged Sumo1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5127)
  TITLE     Addgene's unpublished plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5127)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pJ609 Flag-Sumo1
FEATURES             Location/Qualifiers
     source          1..5127
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      86..674
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     575..594
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     828..845
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     943..960
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     944..960
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1416..1432
                     /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)."
     enhancer        1459..1838
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1839..2042
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     1993..2013
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2087..2106
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2087..2105
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2135..2158
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2503..2520)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2509..2620
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(2738..2758)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2743..3072
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2923..3058
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2985..3004
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3120..3167
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3186..3785
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(3186..3205)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3682..3702
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3915..4036
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3952..3971)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4006..4025
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4089..4880,4881..4949))
                     /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     4712..4731
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4950..5041)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5083..5102
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5108..5127
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 ctcatgacca aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc
       61 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
      121 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
      181 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
      241 tgtagccgta gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc
      301 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
      361 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
      421 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
      481 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
      541 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
      601 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
      661 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
      721 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
      781 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
      841 gcgaggaagc ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc
      901 tgacggatgg cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt
      961 cttaagctcg ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg
     1021 taaaaacccg cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag
     1081 aatatttaag ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg
     1141 agaaaaaagc aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata
     1201 attaaactat tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa
     1261 agagaattaa gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa
     1321 attatacatg tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt
     1381 attatcattt agaataaatt ttgtgtcgcc cttaattgtg agcggataac aattacgagc
     1441 ttcatgcaca gtggcgttga cattgattat tgactagtta ttaatagtaa tcaattacgg
     1501 ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc
     1561 cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca
     1621 tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg
     1681 cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg
     1741 acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt
     1801 ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca
     1861 tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg
     1921 tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact
     1981 ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag
     2041 ctctctggct aactagagaa cccactgctt actggcttat cgaaattaat acgactcact
     2101 ataggggaca cccaagctgg ctagcgacac catggactac aaagacgatg acgacaaggg
     2161 tccaggacct ggtccaggac ctatgtctga ccaggaggca aaaccttcaa ctgaggactt
     2221 gggggataag aaggaaggtg aatatattaa actcaaagtc attggacagg atagcagtga
     2281 gattcacttc aaagtgaaaa tgacaacaca tctcaagaaa ctcaaagaat catactgtca
     2341 aagacagggt gttccaatga attcactcag gtttctcttt gagggtcaga gaattgctga
     2401 taatcatact ccaaaagaac tgggaatgga ggaagaagat gtgattgaag tttatcagga
     2461 acaaacgggg ggtcattcaa cagtttagct cgagaaaatc agcctcgact gtgccttcta
     2521 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     2581 ctcccactgt cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc
     2641 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2701 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2761 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2821 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2881 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2941 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3001 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3061 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     3121 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     3181 aaaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     3241 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     3301 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     3361 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     3421 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     3481 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     3541 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     3601 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     3661 tcctggagac atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     3721 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     3781 cctgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3841 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3901 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3961 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4021 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4081 catggtcatt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4141 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4201 tctggcccca gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca
     4261 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4321 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4381 ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4441 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4501 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4561 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     4621 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     4681 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     4741 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     4801 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     4861 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4921 agggcgacac ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt
     4981 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5041 ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata
     5101 cctgaatatg gctcataaca ccccttg
//
