LOCUS       sequence-379899-        7359 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human wild-type short-form UBASH3A with V5 
            tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7359)
  AUTHORS   Ge Y, Paisie TK, Newman JRB, McIntyre LM, Concannon P
  TITLE     UBASH3A Mediates Risk for Type 1 Diabetes Through Inhibition of 
            T-Cell Receptor-Induced NF-kappaB Signaling.
  JOURNAL   Diabetes. 2017 Jul;66(7):2033-2043. doi: 10.2337/db16-1023. Epub 
            2017 Jun 12.
  PUBMED    28607106
REFERENCE   2  (bases 1 to 7359)
  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"; doi: "10.2337/db16-1023";
            journalName: "Diabetes"; date: "2017-07"; volume: "66"; issue: "7"; 
            pages: "2033-2043"
COMMENT     Sequence Label: V5-WT-short-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7359
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(19..38)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        210..589
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        590..793
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     744..764
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     838..857
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        838..856
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             889..930
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             931..951
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    982..1006
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2882..2906)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2925..2942)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2931..3155
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3201..3629
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3288..3307)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3498..3519
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3638..3658)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3643..3972
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3823..3958
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3885..3904
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4039..4833
                     /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(4093..4112)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4703..4722
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5007..5128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5044..5063)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5098..5117
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5177..5193)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5177..5193)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5190..5212)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5201..5217
                     /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(5225..5231,5232..5249,5250..5255))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5270..5291
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5408..5425)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5579..6167)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5659..5678)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6338..7129,7130..7198))
                     /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     6961..6980
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7199..7303)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatctgct
       61 ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag ctacaacaag
      121 gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt ttgcgctgct
      181 tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt attaatagta
      241 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
      301 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      361 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      421 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      481 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      541 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      601 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      661 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      721 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      781 tataagcaga gctctctggc taactagaga acccactgct tactggctta tcgaaattaa
      841 tacgactcac tatagggaga cccaagctgg ctagttaagc ttaccatggg taagcctatc
      901 cctaaccctc tcctcggtct cgattctacg gaaaacctgt attttcaggg cccggatccg
      961 tcgacgaatt ctgcagatat cacaagtttg tacaaaaaag caggcttcat ggcagcgggg
     1021 gagacgcagc tctacgccaa ggtctccaac aagctcaaga gccgcagcag cccctcgctc
     1081 ctggagcccc tcctggccat gggcttcccg gtgcacaccg cgctgaaagc gttggcagcc
     1141 acggggagga agacggcgga ggaggccttg gcctggctgc atgatcattg caatgaccct
     1201 tccctagacg accccatccc ccaggagtat gcccttttcc tctgtccaac ggggcccctg
     1261 ctggaaaaac ttcaagagtt ctggagagag agcaagcgcc agtgtgcaaa gaacagagct
     1321 catgaggtct tcccacacgt gacactctgt gacttcttca cgtgtgaaga ccagaaggtg
     1381 gaatgcctgt acgaggcgct gaagagagct ggagacaggc tcctgggctc cttccccacg
     1441 gccgtgcctc tggctctcca ctcctccatc agctacctcg gcttcttcgt cagtggcagc
     1501 cccgcagacg tcatccggga attcgccatg accttcgcca cggaagcatc tctcttagca
     1561 gactgctccg tgaagccttg caccaaacag ctgcatctga ccttggccca caagttctac
     1621 ccccaccacc agaggacgct ggagcagctg gccagagcca tccccctggg ccacagctgc
     1681 cagtggaccg cagcactcta ctcccgagac atgcgctttg tgcactacca gaccctgaga
     1741 gccctattcc agtacaaacc ccagaacgtg gatgagctga cgctaagtcc tggtgactac
     1801 atctttgtgg accccacgca gcaggacgaa gccagcgagg gctgggtgat tgggatctca
     1861 cagcggacgg gctgccgggg cttcctgccg gaaaactaca cggatcgagc cagtgagtct
     1921 gacacgtggg tgaagcacag gatgtacacc ttcagtctag ccacagacct gaactccaga
     1981 aaggatggtg aagccagcag cagatgcagc ggggaatttc ttccacaaac ggcaaggagt
     2041 cttagcagct tacaggcctt gcaggctacc gttgcaagga agagtgtgct ggtggttcgc
     2101 cacggggaga gagtggatca gatcttcggg aaggcatggc tgcagcaatg ctccactcct
     2161 gatgggaaat actacaggcc agacctgaat ttcccctgca gtctgcccag acggagtcgt
     2221 gggatcaaag actttgaaaa cgatccccca ttatcatcgt gtggcatttt ccagtccaga
     2281 attgcagggg acgcgctact ggacagtggt atcagaatca gctctgtgtt tgcctcccca
     2341 gccctccgct gtgtgcagac ggccaaactc atcctggaag aactcaaact ggagaaaaaa
     2401 atcaagatac gagtggaacc tggaatcttt gaatggacaa aatgggaagc tggcaagacc
     2461 accccaaccc tcatgagcct ggaagagctg aaagaggcaa atttcaacat tgacactgat
     2521 tacaggcccg cgtttcccct gtccgccctc atgccggccg agagctacca ggagtacatg
     2581 gacaggtgca cggcgagcat ggtgcaaatc gtcaacacct gtccgcagga cacgggtgtc
     2641 atcctaattg tgagtcacgg ctccactctg gactcctgca cgcggccact gctcgggctg
     2701 ccgccccggg aatgtgggga ttttgcccaa ctcgtgagaa agatcccttc cctgggcatg
     2761 tgcttctgtg aagaaaataa agaggaagga aaatgggagt tggtgaaccc accggtgaag
     2821 accctgaccc acggggcgaa cgcagcattt aactggagga actggatctc aggcaactga
     2881 aacccagctt tcttgtacaa agtggtgata aacccgctga tcagcctcga ctgtgccttc
     2941 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3001 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3061 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3121 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     3181 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     3241 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     3301 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     3361 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     3421 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     3481 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     3541 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     3601 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     3661 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     3721 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3781 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3841 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3901 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3961 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     4021 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     4081 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     4141 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     4201 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     4261 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     4321 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     4381 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     4441 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     4501 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     4561 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     4621 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     4681 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     4741 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     4801 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     4861 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     4921 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     4981 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     5041 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     5101 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     5161 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     5221 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     5281 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     5341 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     5401 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     5461 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     5521 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     5581 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     5641 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     5701 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     5761 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     5821 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     5881 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     5941 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     6001 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     6061 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     6121 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     6181 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     6241 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     6301 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     6361 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     6421 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     6481 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     6541 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     6601 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     6661 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     6721 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     6781 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     6841 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     6901 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     6961 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     7021 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     7081 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     7141 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     7201 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     7261 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     7321 tgccacctga cgtcgacgga tcgggagatc tcccgatcc
//
