LOCUS       sequence-14175-e        5897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5897)
  AUTHORS   Yang W, Paschen W
  TITLE     Gene expression and cell growth are modified by silencing SUMO2 and 
            SUMO3 expression.
  JOURNAL   Biochem Biophys Res Commun. 2009 Apr 24. 382(1):215-8.
  PUBMED    19275883
REFERENCE   2  (bases 1 to 5897)
  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: "Biochem 
            Biophys Res Commun. 2009 Apr 24. 382(1):215-8."
COMMENT     Sequence Label: pcDNA6.2-GW/EmGFP-miR-SUMO23
FEATURES             Location/Qualifiers
     source          1..5897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        384..587
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     538..558
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     promoter        632..650
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    680..704
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(713..715,716..718,719..1432)
                     /codon_start=1
                     /product="Emerald GFP"
                     /label=EmGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHKVYITADKQKNGIK
                     VNFKTRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(758..779)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1019..1038)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1366..1387
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     ncRNA           1492..1518
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           1582..1623
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           1630..1656
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           1720..1761
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     protein_bind    complement(1790..1814)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(1856..1875)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1900..1948
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        complement(2119..2137)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2142..2158)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2142..2158)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     rep_origin      2226..2654
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2313..2332)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2523..2544
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2663..2683)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2668..2997
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2848..2983
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2910..2929
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3045..3092
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3111..3509
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    3667..3788
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3704..3723)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3758..3777
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3823..3840)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3994..4582)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4074..4093)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(4678..5469)
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
ORIGIN
        1 gttgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat gggcgtggat
      421 agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat gggagtttgt
      481 tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc ccattgacgc
      541 aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc tggctaacta
      601 gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg gagtcccaag
      661 ctggctagtt aagctatcaa caagtttgta caaaaaagca ggctttaaaa ccatggtgag
      721 caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt
      781 aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct
      841 gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac
      901 caccttcacc tacggcgtgc agtgcttcgc ccgctacccc gaccacatga agcagcacga
      961 cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga
     1021 cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg
     1081 catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga
     1141 gtacaactac aacagccaca aggtctatat caccgccgac aagcagaaga acggcatcaa
     1201 ggtgaacttc aagacccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta
     1261 ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag
     1321 cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga
     1381 gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt aagctaagca
     1441 cttcgtggcc gtcgatcgtt taaagggagg tagtgagtcg accagtggat cctggaggct
     1501 tgctgaaggc tgtatgctga atcgaatctg cctcattgac gttttggcca ctgactgacg
     1561 tcaatgacag attcgattca ggacacaagg cctgttacta gcactcacat ggaacaaatg
     1621 gcccagatcc tggaggcttg ctgaaggctg tatgctgtga tctgcctcat tgacaaacgt
     1681 tttggccact gactgacgtt tgtcagaggc agatcacagg acacaaggcc tgttactagc
     1741 actcacatgg aacaaatggc ccagatctgg ccgcactcga gatatctaga cccagctttc
     1801 ttgtacaaag tggttgatct agagggcccg cggttcgctg atgggggagg ctaactgaaa
     1861 cacggaagga gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata
     1921 aaacgcacgg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca gggctggcac
     1981 tctgtcgata ccccaccgtg accccattgg ggccaatacg cccgcgtttc ttccttttcc
     2041 ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt cggggcggca
     2101 ggccctgcca tagcatcccc tatagtgagt cgtattacat ggtcatagct gtttcctggc
     2161 agctctggcc cgtgtctcaa aatctctgat ggatctgcgc agctggggct ctagggggta
     2221 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     2281 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     2341 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     2401 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     2461 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     2521 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     2581 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     2641 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     2701 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     2761 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2821 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     2881 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     2941 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     3001 ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac aattaatcat
     3061 cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc atggccaagc
     3121 ctttgtctca agaagaatcc accctcattg aaagagcaac ggctacaatc aacagcatcc
     3181 ccatctctga agactacagc gtcgccagcg cagctctctc tagcgacggc cgcatcttca
     3241 ctggtgtcaa tgtatatcat tttactgggg gaccttgtgc agaactcgtg gtgctgggca
     3301 ctgctgctgc tgcggcagct ggcaacctga cttgtatcgt cgcgatcgga aatgagaaca
     3361 ggggcatctt gagcccctgc ggacggtgcc gacaggtgct tctcgatctg catcctggga
     3421 tcaaagccat agtgaaggac agtgatggac agccgacggc agttgggatt cgtgaattgc
     3481 tgccctctgg ttatgtgtgg gagggctaag cacttcgtgg ccgaggagca ggactgacac
     3541 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3601 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3661 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3721 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3781 gtatcttatc atgtctgtat accgtcgctc ttccgctgct tcctcgctca ctgactcgct
     3841 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     3901 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     3961 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     4021 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     4081 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     4141 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     4201 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     4261 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     4321 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     4381 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     4441 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     4501 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     4561 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     4621 gtggaacgac gcgtaactca cgttaaggga ttttggtcat gggtggctcg acgagggtta
     4681 tttgccgact accttggtga tctcgccttt cacgtagtgg acaaattctt ccaactgatc
     4741 tgcgcgcgag gccaagcgat cttcttcttg tccaagataa gcctgtctag cttcaagtat
     4801 gacgggctga tactgggccg gcaggcgctc cattgcccag tcggcagcga catccttcgg
     4861 cgcgattttg ccggttactg cgctgtacca aatgcgggac aacgtaagca ctacatttcg
     4921 ctcatcgcca gcccagtcgg gcggcgagtt ccatagcgtt aaggtttcat ttagcgcctc
     4981 aaatagatcc tgttcaggaa ccggatcaaa gagttcctcc gccgctggac ctaccaaggc
     5041 aacgctatgt tctcttgctt ttgtcagcaa gatagccaga tcaatgtcga tcgtggctgg
     5101 ctcgaagata cctgcaagaa tgtcattgcg ctgccattct ccaaattgca gttcgcgctt
     5161 agctggataa cgccacggaa tgatgtcgtc gtgcacaaca atggtgactt ctacagcgcg
     5221 gagaatctcg ctctctccag gggaagccga agtttccaaa aggtcgttga tcaaagctcg
     5281 ccgcgttgtt tcatcaagcc ttacggtcac cgtaaccagc aaatcaatat cactgtgtgg
     5341 cttcaggccg ccatccactg cggagccgta caaatgtacg gccagcaacg tcggttcgag
     5401 atggcgctcg atgacgccaa ctacctctga tagttgagtc gatacttcgg cgatcaccgc
     5461 ttccctcata atgtttaact ttgttttagg gcgactgccc tgctgcgtaa catcgttgct
     5521 gctccataac atcaaacatc gacccacggc gtaacgcgct tgctgcttgg atgcccgagg
     5581 catagactgt accccaaaaa aacagtcata acaagccatg aaaaccgcca ctgcgccgtt
     5641 accaccgctg cgttcggtca aggttctgga ccagttgcgt gagcgcatac gctacttgca
     5701 ttacagctta cgaaccgaac aggcttatgt ccactgggtt cgtgccttca tccgtttcca
     5761 cggtgtgcgt cacccggcaa ccttgggtag cagcgaagtc gaggcatttc tgtcctggct
     5821 ggtctagaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     5881 acgggccaga tatacgc
//
