LOCUS       sequence-379898-        7473 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human wild-type long-form UBASH3A with V5 
            tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7473)
  TITLE     Concannon Lab UBASH3A Plasmids 2022
REFERENCE   2  (bases 1 to 7473)
  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"
COMMENT     Sequence Label: V5-WT-long-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7473
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        178..557
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        558..761
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     712..732
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     806..825
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        806..824
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             857..898
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             899..919
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    950..974
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2964..2988)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(3007..3024)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3013..3237
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3283..3711
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3370..3389)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3580..3601
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3720..3740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3725..4054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3905..4040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3967..3986
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4121..4915
                     /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(4175..4194)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4785..4804
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5089..5210
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5126..5145)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5180..5199
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5259..5275)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5259..5275)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5272..5294)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5283..5299
                     /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(5307..5313,5314..5331,5332..5337))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5352..5373
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5490..5507)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5661..6249)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5741..5760)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6420..7211,7212..7280))
                     /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     7043..7062
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7281..7385)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(join(7460..7473,1..6))
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
       61 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
      121 tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgttgac
      181 attgattatt gactagttat taatagtaat caattacggg gtcattagtt catagcccat
      241 atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga ccgcccaacg
      301 acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca atagggactt
      361 tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca gtacatcaag
      421 tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg cccgcctggc
      481 attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc tacgtattag
      541 tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt ggatagcggt
      601 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      661 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      721 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      781 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      841 agttaagctt accatgggta agcctatccc taaccctctc ctcggtctcg attctacgga
      901 aaacctgtat tttcagggcc cggatccgtc gacgaattct gcagatatca caagtttgta
      961 caaaaaagca ggcttcatgg cagcggggga gacgcagctc tacgccaagg tctccaacaa
     1021 gctcaagagc cgcagcagcc cctcgctcct ggagcccctc ctggccatgg gcttcccggt
     1081 gcacaccgcg ctgaaagcgt tggcagccac ggggaggaag acggcggagg aggccttggc
     1141 ctggctgcat gatcattgca atgacccttc cctagacgac cccatccccc aggagtatgc
     1201 ccttttcctc tgtccaacgg ggcccctgct ggaaaaactt caagagttct ggagagagag
     1261 caagcgccag tgtgcaaaga acagagctca tgaggtcttc ccacacgtga cactctgtga
     1321 cttcttcacg tgtgaagacc agaaggtgga atgcctgtac gaggcgctga agagagctgg
     1381 agacaggctc ctgggctcct tccccacggc cgtgcctctg gctctccact cctccatcag
     1441 ctacctcggc ttcttcgtca gtggcagccc cgcagacgtc atccgggaat tcgccatgac
     1501 cttcgccacg gaagcatctc tcttagcagg cacttccgtt tcccgcttct ggattttcag
     1561 ccaggtgcct ggacatggcc ctaacctgag gctgagcaat ttaactagag cctccttcgt
     1621 gagccactac atccttcaaa aatactgctc cgtgaagcct tgcaccaaac agctgcatct
     1681 gaccttggcc cacaagttct acccccacca ccagaggacg ctggagcagc tggccagagc
     1741 catccccctg ggccacagct gccagtggac cgcagcactc tactcccgag acatgcgctt
     1801 tgtgcactac cagaccctga gagccctatt ccagtacaaa ccccagaacg tggatgagct
     1861 gacgctaagt cctggtgact acatctttgt ggaccccacg cagcaggacg aagccagcga
     1921 gggctgggtg attgggatct cacagcggac gggctgccgg ggcttcctgc cggaaaacta
     1981 cacggatcga gccagtgagt ctgacacgtg ggtgaagcac aggatgtaca ccttcagtct
     2041 agccacagac ctgaactcca gaaaggatgg tgaagccagc agcagatgca gcggggaatt
     2101 tcttccacaa acggcaagga gtcttagcag cttacaggcc ttgcaggcta ccgttgcaag
     2161 gaagagtgtg ctggtggttc gccacgggga gagagtggat cagatcttcg ggaaggcatg
     2221 gctgcagcaa tgctccactc ctgatgggaa atactacagg ccagacctga atttcccctg
     2281 cagtctgccc aggcggagtc gtgggatcaa agactttgaa aacgatcccc cattatcatc
     2341 gtgtggcatt ttccagtcca gaattgcagg ggacgcgcta ctggacagtg gtatcagaat
     2401 cagctctgtg tttgcctccc cagccctccg ctgtgtgcag acggccaaac tcatcctgga
     2461 agaactcaaa ctggagaaaa aaatcaagat acgagtggaa cctggaatct ttgaatggac
     2521 aaaatgggaa gctggcaaaa ccaccccaac cctcatgagc ctggaagagc tgaaagaggc
     2581 aaatttcaac attgacactg attacaggcc cgcgtttccc ctgtccgccc tcatgccggc
     2641 cgagagctac caggagtaca tggacaggtg cacggcgagc atggtgcaaa tcgtcaacac
     2701 ctgcccgcag gacacgggtg tcatcctaat tgtgagtcac ggctccactc tggactcctg
     2761 cacgcggcca ctgctcgggc tgccgccccg ggaatgtggg gattttgccc aactcgtgag
     2821 aaagatccct tccctgggca tgtgcttctg tgaagaaaat aaagaggaag gaaaatggga
     2881 gttggtgaac ccaccggtga agaccctgac ccacggggcg aacgcagcat ttaactggag
     2941 gaactggatc tcaggcaact gaaacccagc tttcttgtac aaagtggtga taaacccgct
     3001 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     3061 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     3121 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3181 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3241 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3301 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3361 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3421 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3481 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3541 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3601 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3661 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3721 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3781 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3841 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3901 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3961 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4021 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4081 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4141 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4201 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4261 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4321 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4381 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4441 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4501 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4561 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4621 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4681 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4741 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4801 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4861 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4921 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4981 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     5041 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     5101 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     5161 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5221 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5281 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5341 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5401 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5461 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5521 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5581 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5641 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5701 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5761 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5821 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5881 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5941 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6001 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6061 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6121 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6181 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6241 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6301 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6361 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6421 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6481 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6541 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6601 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6661 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6721 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6781 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6841 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6901 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6961 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7021 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7081 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7141 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7201 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7261 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7321 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7381 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     7441 ccctatggtg cactctcagt acaatctgct ctg
//
