LOCUS       sequence-386311-        7544 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human short-form UBASH3A lacking the UBA 
            domain with V5-6xHis tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7544)
  TITLE     Concannon Lab Plasmids
REFERENCE   2  (bases 1 to 7544)
  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: deltaUBA-short-UBASH3A-V5-6xHis
FEATURES             Location/Qualifiers
     source          1..7544
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(166..185)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        473..1651
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          704..1642
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1599..1619
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1668..1687
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1668..1686
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1718..1742
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3492..3516)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3542..3583
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3593..3610
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3633..3650)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3639..3863
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3909..4337
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3996..4015)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4206..4227
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4346..4366)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4351..4680
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4531..4666
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4593..4612
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4728..4775
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4794..5192
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5350..5471
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5387..5406)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5441..5460
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5520..5536)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5520..5536)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5533..5555)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5544..5560
                     /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(5568..5574,5575..5592,5593..5598))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5613..5634
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5751..5768)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5911..6499)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5991..6010)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6670..7461,7462..7530))
                     /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     7293..7312
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7531..7544,1..91))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
       61 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
      121 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
      181 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      241 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      301 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      361 ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt tttaaacaga
      421 gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa ttggctccgg
      481 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
      541 cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
      601 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
      661 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg ccgtgtgtgg
      721 ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa ttacttccac
      781 ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt gggagagttc
      841 gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct ggcctgggcg
      901 ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg ctttcgataa
      961 gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct ggcaagatag
     1021 tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg ccgcgggcgg
     1081 cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc gagcgcggcc
     1141 accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc ctggcctcgc
     1201 gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac cagttgcgtg
     1261 agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga ggacgcggcg
     1321 ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc cgtcctcagc
     1381 cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg attagttctc
     1441 gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga tggagtttcc
     1501 ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt aattctcctt
     1561 ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga cagtggttca
     1621 aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtactaa tacgactcac
     1681 tatagggaga cccaagctgg ctaggtaagc ttgatcaaca agtttgtaca aaaaagcagg
     1741 cttcatggca gcgggggaga cgcagctcta cgccaaggtc tccaacaagc tcaagagccg
     1801 ctgcaatgac ccttccctag acgaccccat cccccaggag tatgcccttt tcctctgtcc
     1861 aacggggccc ctgctggaaa aacttcaaga gttctggaga gagagcaagc gccagtgtgc
     1921 aaagaacaga gctcatgagg tcttcccaca cgtgacactc tgtgacttct tcacgtgtga
     1981 agaccagaag gtggaatgcc tgtacgaggc gctgaagaga gctggagaca ggctcctggg
     2041 ctccttcccc acggccgtgc ctctggctct ccactcctcc atcagctacc tcggcttctt
     2101 cgtcagtggc agccccgcag acgtcatccg ggaattcgcc atgaccttcg ccacggaagc
     2161 atctctctta gcagactgct ccgtgaagcc ttgcaccaaa cagctgcatc tgaccttggc
     2221 ccacaagttc tacccccacc accagaggac gctggagcag ctggccagag ccatccccct
     2281 gggccacagc tgccagtgga ccgcagcact ctactcccga gacatgcgct ttgtgcacta
     2341 ccagaccctg agagccctat tccagtacaa accccagaac gtggatgagc tgacgctaag
     2401 tcctggtgac tacatctttg tggaccccac gcagcaggac gaagccagcg agggctgggt
     2461 gattgggatc tcacagcgga cgggctgccg gggcttcctg ccggaaaact acacggatcg
     2521 agccagtgag tctgacacgt gggtgaagca caggatgtac accttcagtc tagccacaga
     2581 cctgaactcc agaaaggatg gtgaagccag cagcagatgc agcggggaat ttcttccaca
     2641 aacggcaagg agtcttagca gcttacaggc cttgcaggct accgttgcaa ggaagagtgt
     2701 gctggtggtt cgccacgggg agagagtgga tcagatcttc gggaaggcat ggctgcagca
     2761 atgctccact cctgatggga aatactacag gccagacctg aatttcccct gcagtctgcc
     2821 cagacggagt cgtgggatca aagactttga aaacgatccc ccattatcat cgtgtggcat
     2881 tttccagtcc agaattgcag gggacgcgct actggacagt ggtatcagaa tcagctctgt
     2941 gtttgcctcc ccagccctcc gctgtgtgca gacggccaaa ctcatcctgg aagaactcaa
     3001 actggagaaa aaaatcaaga tacgagtgga acctggaatc tttgaatgga caaaatggga
     3061 agctggcaag accaccccaa ccctcatgag cctggaagag ctgaaagagg caaatttcaa
     3121 cattgacact gattacaggc ccgcgtttcc cctgtccgcc ctcatgccgg ccgagagcta
     3181 ccaggagtac atggacaggt gcacggcgag catggtgcaa atcgtcaaca cctgtccgca
     3241 ggacacgggt gtcatcctaa ttgtgagtca cggctccact ctggactcct gcacgcggcc
     3301 actgctcggg ctgccgcccc gggaatgtgg ggattttgcc caactcgtga gaaagatccc
     3361 ttccctgggc atgtgcttct gtgaagaaaa taaagaggaa ggaaaatggg agttggtgaa
     3421 cccaccggtg aagaccctga cccacggggc gaacgcagca tttaactgga ggaactggat
     3481 ctcaggcaac aacccagctt tcttgtacaa agtggttgat ctagagggcc cgcggttcga
     3541 aggtaagcct atccctaacc ctctcctcgg tctcgattct acgcgtaccg gtcatcatca
     3601 ccatcaccat tgagtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc
     3661 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     3721 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     3781 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     3841 tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg
     3901 gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     3961 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4021 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4081 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     4141 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4201 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     4261 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     4321 aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca
     4381 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt
     4441 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     4501 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
     4561 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct
     4621 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
     4681 aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt gacaattaat
     4741 catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa accatggcca
     4801 agcctttgtc tcaagaagaa tccaccctca ttgaaagagc aacggctaca atcaacagca
     4861 tccccatctc tgaagactac agcgtcgcca gcgcagctct ctctagcgac ggccgcatct
     4921 tcactggtgt caatgtatat cattttactg ggggaccttg tgcagaactc gtggtgctgg
     4981 gcactgctgc tgctgcggca gctggcaacc tgacttgtat cgtcgcgatc ggaaatgaga
     5041 acaggggcat cttgagcccc tgcggacggt gccgacaggt gcttctcgat ctgcatcctg
     5101 ggatcaaagc catagtgaag gacagtgatg gacagccgac ggcagttggg attcgtgaat
     5161 tgctgccctc tggttatgtg tgggagggct aagcacttcg tggccgagga gcaggactga
     5221 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5281 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5341 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     5401 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     5461 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     5521 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     5581 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     5641 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     5701 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     5761 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5821 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5881 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     5941 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     6001 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     6061 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     6121 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     6181 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     6241 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     6301 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt
     6361 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     6421 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     6481 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     6541 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     6601 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     6661 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     6721 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     6781 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     6841 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     6901 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     6961 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     7021 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     7081 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     7141 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     7201 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     7261 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     7321 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     7381 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     7441 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     7501 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttc
//
