LOCUS       sequence-386309-        7236 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human short-form UBASH3A lacking the UBA 
            domain with V5 tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7236)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 7236)
  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-deltaUBA-short-UBASH3A
FEATURES             Location/Qualifiers
     source          1..7236
                     /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             914..955
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             956..976
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     protein_bind    1007..1031
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(2784..2808)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2827..2844)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2833..3057
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3103..3531
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3190..3209)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3400..3421
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3540..3560)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3545..3874
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3725..3860
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3787..3806
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3941..4735
                     /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(3995..4014)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4605..4624
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4909..5030
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4946..4965)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5000..5019
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5079..5095)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5079..5095)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5092..5114)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5103..5119
                     /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(5127..5133,5134..5151,5152..5157))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5172..5193
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5310..5327)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5481..6069)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5561..5580)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6240..7031,7032..7100))
                     /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     6863..6882
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7101..7205)
                     /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 gctggctagt
      901 taagcttacc atgggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacggaaaa
      961 cctgtatttt cagggcccgg atccgtcgac gaattctgca gatatcacaa gtttgtacaa
     1021 aaaagcaggc ttcatggcag cgggggagac gcagctctac gccaaggtct ccaacaagct
     1081 caagagccgc tgcaatgacc cttccctaga cgaccccatc ccccaggagt atgccctttt
     1141 cctctgtcca acggggcccc tgctggaaaa acttcaagag ttctggagag agagcaagcg
     1201 ccagtgtgca aagaacagag ctcatgaggt cttcccacac gtgacactct gtgacttctt
     1261 cacgtgtgaa gaccagaagg tggaatgcct gtacgaggcg ctgaagagag ctggagacag
     1321 gctcctgggc tccttcccca cggccgtgcc tctggctctc cactcctcca tcagctacct
     1381 cggcttcttc gtcagtggca gccccgcaga cgtcatccgg gaattcgcca tgaccttcgc
     1441 cacggaagca tctctcttag cagactgctc cgtgaagcct tgcaccaaac agctgcatct
     1501 gaccttggcc cacaagttct acccccacca ccagaggacg ctggagcagc tggccagagc
     1561 catccccctg ggccacagct gccagtggac cgcagcactc tactcccgag acatgcgctt
     1621 tgtgcactac cagaccctga gagccctatt ccagtacaaa ccccagaacg tggatgagct
     1681 gacgctaagt cctggtgact acatctttgt ggaccccacg cagcaggacg aagccagcga
     1741 gggctgggtg attgggatct cacagcggac gggctgccgg ggcttcctgc cggaaaacta
     1801 cacggatcga gccagtgagt ctgacacgtg ggtgaagcac aggatgtaca ccttcagtct
     1861 agccacagac ctgaactcca gaaaggatgg tgaagccagc agcagatgca gcggggaatt
     1921 tcttccacaa acggcaagga gtcttagcag cttacaggcc ttgcaggcta ccgttgcaag
     1981 gaagagtgtg ctggtggttc gccacgggga gagagtggat cagatcttcg ggaaggcatg
     2041 gctgcagcaa tgctccactc ctgatgggaa atactacagg ccagacctga atttcccctg
     2101 cagtctgccc agacggagtc gtgggatcaa agactttgaa aacgatcccc cattatcatc
     2161 gtgtggcatt ttccagtcca gaattgcagg ggacgcgcta ctggacagtg gtatcagaat
     2221 cagctctgtg tttgcctccc cagccctccg ctgtgtgcag acggccaaac tcatcctgga
     2281 agaactcaaa ctggagaaaa aaatcaagat acgagtggaa cctggaatct ttgaatggac
     2341 aaaatgggaa gctggcaaga ccaccccaac cctcatgagc ctggaagagc tgaaagaggc
     2401 aaatttcaac attgacactg attacaggcc cgcgtttccc ctgtccgccc tcatgccggc
     2461 cgagagctac caggagtaca tggacaggtg cacggcgagc atggtgcaaa tcgtcaacac
     2521 ctgtccgcag gacacgggtg tcatcctaat tgtgagtcac ggctccactc tggactcctg
     2581 cacgcggcca ctgctcgggc tgccgccccg ggaatgtggg gattttgccc aactcgtgag
     2641 aaagatccct tccctgggca tgtgcttctg tgaagaaaat aaagaggaag gaaaatggga
     2701 gttggtgaac ccaccggtga agaccctgac ccacggggcg aacgcagcat ttaactggag
     2761 gaactggatc tcaggcaact gaaacccagc tttcttgtac aaagtggtga taaacccgct
     2821 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     2881 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     2941 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     3001 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     3061 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3121 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3181 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3241 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     3301 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3361 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3421 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     3481 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3541 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3601 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3661 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     3721 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     3781 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     3841 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     3901 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     3961 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4021 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4081 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4141 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4201 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4261 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4321 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4381 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4441 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4501 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4561 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4621 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4681 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     4741 actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat
     4801 tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg
     4861 atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt
     4921 gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt
     4981 ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt
     5041 ataccgtcga cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga
     5101 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     5161 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     5221 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     5281 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5341 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5401 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5461 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     5521 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     5581 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     5641 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     5701 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     5761 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     5821 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     5881 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     5941 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6001 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6061 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6121 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6181 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6241 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6301 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6361 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6421 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6481 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     6541 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     6601 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     6661 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     6721 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     6781 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     6841 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     6901 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     6961 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7021 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7081 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7141 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7201 ccgcgcacat ttccccgaaa agtgccacct gacgtc
//
