LOCUS       sequence-379904-        7781 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of human wild-type long-form UBASH3A with 
            V5-6xHis tag .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7781)
  TITLE     Concannon Lab UBASH3A Plasmids 2022
REFERENCE   2  (bases 1 to 7781)
  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: WT-long-UBASH3A-V5-6xHis
FEATURES             Location/Qualifiers
     source          1..7781
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        237..1415
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          468..1406
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1363..1383
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     1432..1451
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1432..1450
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1482..1506
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(3493..3517)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3543..3584
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             3594..3611
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3634..3651)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3640..3864
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3910..4338
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3997..4016)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4207..4228
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4347..4367)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4352..4681
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4532..4667
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4594..4613
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4729..4776
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4795..5193
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /product="blasticidin S deaminase"
                     /label=BSD
                     /note="confers resistance to blasticidin"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     polyA_signal    5351..5472
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5388..5407)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5442..5461
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5521..5537)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5521..5537)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5534..5556)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5545..5561
                     /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(5569..5575,5576..5593,5594..5599))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5614..5635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5752..5769)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5912..6500)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5992..6011)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6671..7462,7463..7531))
                     /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     7294..7313
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7532..7636)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7711..7730)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat tgcatgaaga
       61 atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga tatacgcgtt
      121 gacattgatt attgactagg cttttgcaaa aagctttgca aagatggata aagttttaaa
      181 cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg ggaattggct
      241 ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag ttggggggag
      301 gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg gaaagtgatg
      361 tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata agtgcagtag
      421 tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta agtgccgtgt
      481 gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct tgaattactt
      541 ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt gggtgggaga
      601 gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag gcctggcctg
      661 ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc gctgctttcg
      721 ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt ttctggcaag
      781 atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt ggggccgcgg
      841 gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc ctgcgagcgc
      901 ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg gtgcctggcc
      961 tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg gcaccagttg
     1021 cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa tggaggacgc
     1081 ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc tttccgtcct
     1141 cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac ctcgattagt
     1201 tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat gcgatggagt
     1261 ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg atgtaattct
     1321 ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct cagacagtgg
     1381 ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat tagcttggta ctaatacgac
     1441 tcactatagg gagacccaag ctggctaggt aagcttgatc aacaagtttg tacaaaaaag
     1501 caggcttcat ggcagcgggg gagacgcagc tctacgccaa ggtctccaac aagctcaaga
     1561 gccgcagcag cccctcgctt ctggagcccc tcctggccat gggcttcccg gtgcacaccg
     1621 cgctgaaagc gttggcagcc acggggagga agacggcgga ggaggccttg gcctggctgc
     1681 atgatcattg caatgaccct tccctagacg accccatccc ccaggagtat gcccttttcc
     1741 tctgtccaac ggggcccctg ctggaaaaac ttcaagagtt ctggagagag agcaagcgcc
     1801 agtgtgcaaa gaacagagct catgaggtct tcccacacgt gacactctgt gacttcttca
     1861 cgtgtgaaga ccagaaggtg gaatgcctgt acgaggcgct gaagagagct ggagacaggc
     1921 tcctgggctc cttccccacg gccgtgcctc tggctctcca ctcctccatc agctacctcg
     1981 gcttcttcgt cagtggcagc cccgcagacg tcatccggga attcgccatg accttcgcca
     2041 cggaagcatc tctcttagca ggcacttccg tttcccgctt ctggattttc agccaggtgc
     2101 ctggacatgg ccctaacctg aggctgagca atttaactag agcctccttc gtgagccact
     2161 acatccttca aaaatactgc tccgtgaagc cttgcaccaa acagctgcat ctgaccttgg
     2221 cccacaagtt ctacccccac caccagagga cgctggagca gctggccaga gccatccccc
     2281 tgggccacag ctgccagtgg accgcagcac tctactcccg agacatgcgc tttgtgcact
     2341 accagaccct gagagcccta ttccagtaca aaccccagaa cgtggatgag ctgacgctaa
     2401 gtcctggtga ctacatcttt gtggacccca cgcagcagga cgaagccagc gagggctggg
     2461 tgattgggat ctcacagcgg acgggctgcc ggggcttcct gccggaaaac tacacggatc
     2521 gagccagtga gtctgacacg tgggtgaagc acaggatgta caccttcagt ctagccacag
     2581 acctgaactc cagaaaggat ggtgaagcca gcagcagatg cagcggggaa tttcttccac
     2641 aaacggcaag gagtcttagc agcttacagg ccttgcaggc taccgttgca aggaagagtg
     2701 tgctggtggt tcgccacggg gagagagtgg atcagatctt cgggaaggca tggctgcagc
     2761 aatgctccac tcctgatggg aaatactaca ggccagacct gaatttcccc tgcagtctgc
     2821 ccaggcggag tcgtgggatc aaagactttg aaaacgatcc cccattatca tcgtgtggca
     2881 ttttccagtc cagaattgca ggggacgcgc tactggacag tggtatcaga atcagctctg
     2941 tgtttgcctc cccagccctc cgctgtgtgc agacggccaa actcatcctg gaagaactca
     3001 aactggagaa aaaaatcaag atacgagtgg aacctggaat ctttgaatgg acaaaatggg
     3061 aagctggcaa aaccacccca accctcatga gcctggaaga gctgaaagag gcaaatttca
     3121 acattgacac tgattacagg cccgcgtttc ccctgtccgc cctcatgccg gccgagagct
     3181 accaggagta catggacagg tgcacggcga gcatggtgca aatcgtcaac acctgcccgc
     3241 aggacacggg tgtcatccta attgtgagtc acggctccac tctggactcc tgcacgcggc
     3301 cactgctcgg gctgccgccc cgggaatgtg gggattttgc ccaactcgtg agaaagatcc
     3361 cttccctggg catgtgcttc tgtgaagaaa ataaagagga aggaaaatgg gagttggtga
     3421 acccaccggt gaagaccctg acccacgggg cgaacgcagc atttaactgg aggaactgga
     3481 tctcaggcaa caacccagct ttcttgtaca aagtggttga tctagagggc ccgcggttcg
     3541 aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc
     3601 accatcacca ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc
     3661 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3721 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3781 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3841 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3901 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3961 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     4021 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     4081 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     4141 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     4201 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     4261 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     4321 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     4381 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     4441 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     4501 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     4561 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     4621 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     4681 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     4741 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatggcc
     4801 aagcctttgt ctcaagaaga atccaccctc attgaaagag caacggctac aatcaacagc
     4861 atccccatct ctgaagacta cagcgtcgcc agcgcagctc tctctagcga cggccgcatc
     4921 ttcactggtg tcaatgtata tcattttact gggggacctt gtgcagaact cgtggtgctg
     4981 ggcactgctg ctgctgcggc agctggcaac ctgacttgta tcgtcgcgat cggaaatgag
     5041 aacaggggca tcttgagccc ctgcggacgg tgccgacagg tgcttctcga tctgcatcct
     5101 gggatcaaag ccatagtgaa ggacagtgat ggacagccga cggcagttgg gattcgtgaa
     5161 ttgctgccct ctggttatgt gtgggagggc taagcacttc gtggccgagg agcaggactg
     5221 acacgtgcta cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat
     5281 cgttttccgg gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt
     5341 cgcccacccc aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     5401 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     5461 caatgtatct tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg
     5521 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5581 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5641 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     5701 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     5761 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     5821 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     5881 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     5941 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6001 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6061 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     6121 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     6181 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     6241 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     6301 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     6361 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     6421 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     6481 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     6541 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     6601 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     6661 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     6721 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     6781 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     6841 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     6901 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     6961 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     7021 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     7081 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     7141 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     7201 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     7261 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     7321 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     7381 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     7441 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     7501 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     7561 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     7621 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag
     7681 atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag
     7741 ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag t
//
