LOCUS       sequence-311860-        9304 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for mammalian overexpression of truncated (delta1-536) ER isoform of
            rat USP19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9304)
  TITLE     rat USP19 constructs
REFERENCE   2  (bases 1 to 9304)
  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: pcDNA-HisFlag-rUSP19ER deltaN1
FEATURES             Location/Qualifiers
     source          1..9304
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        106..485
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        486..689
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     640..660
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     734..753
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        734..752
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      859..868
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             871..888
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             889..954
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(4764..4781)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4770..4994
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5040..5468
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5127..5146)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5337..5358
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5477..5497)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5482..5811
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5662..5797
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5878..6672
                     /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(5932..5951)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6542..6561
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    6848..6969
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6885..6904)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6939..6958
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7018..7034)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7018..7034)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7031..7053)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7042..7058
                     /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(7066..7072,7073..7090,7091..7096))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7111..7132
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7249..7266)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(7420..8008)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(7500..7519)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(8179..8970,8971..9039))
                     /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     8802..8821
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9040..9144)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(9219..9238)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
       61 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
      121 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
      181 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
      241 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
      301 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
      361 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
      421 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
      481 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
      541 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
      601 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
      661 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
      721 ggcttatcga aattaatacg actcactata gggagaccca agctggctag ttaagcttgg
      781 taccgagctc ggatccacta gtccagtgtg gtggaattct gcagatatcc agcacagtgg
      841 cggccgctcg agtctagagc caccatggca catcatcatc atcatcacga ctacaaagac
      901 catgacggtg attataaaga tcatgacatc gattacaagg atgacgatga caagcgcccg
      961 ggctgcagga attcagctcg aaaccagaag agggcccagg gccgtggtga ggtaggctca
     1021 ggggctagcc ctggggcaca ggcagggccc agtgccaaga gggctgttca tctctgcaga
     1081 gggccagaag gtgaagggtc catggatggg cctggccccc agggtgatgc cccatccttc
     1141 ctgtccgact cagctacgca ggttgaggct gaggagcagc tccatgttcc accagtgaac
     1201 cctcaaacaa gtctcttggg ctcagagaag aatttagccc ttttgacagt agagaagaca
     1261 gtgtccccca ggagtgactc agtctcccca gtcatgatcc ggaacagaga ccctgagaaa
     1321 gatgaccatt tcaaagagga gatggcagtt ggagctgatc ctgcagcctt agcggatgaa
     1381 ccagagtcta tggtgaacct ggcatttgtc aagaatgatt cgtatgagaa ggggccggat
     1441 tcagtggtgg tgcacgtgta tgtgaaggag agccgcaggg atacctctcg agtacttttc
     1501 cgagagcagg acttcacgct gatcttccag accagggatg gaaattttct gaggctgcat
     1561 ccgggctgtg ggccccacac catcttccgc tggcaggtga agctcaggaa cttgattgag
     1621 ccagagcaat gtacgttttg tttcacggcc tctcgaatcg atatctgcct ccggaagcgg
     1681 cagagtcagc gctggggggg actggaggcc cctgctacac gagtgggtgg tgcaaaggtt
     1741 gccgtgccga caggtccaac ccctttggat tcaacccctc caggaggtgg ccccctccct
     1801 ctgactggcc aggaggaagc cagggctgta gagaaggaaa aacccaaggc tcggtcagag
     1861 gactcggggc tggatggtgt ggtggctcgc acccccttgg agcatgttac cccaaagcca
     1921 gaaccgcact tggcctcgcc caaacccaca tgtatggtgc ctccaatgcc tcatagtcca
     1981 gtgagtggag atagtgtgga ggaggatgaa gaggaagaga agaaggtttg tctcccaggc
     2041 tttactggcc tcgtcaactt agggaacacc tgcttcatga atagtgtcat tcagtctctg
     2101 tccaacactc gggagcttcg tgacttcttc cacgaccgat cctttgaggc tgagattaac
     2161 tacaataacc cattggggac tggtgggcgt ctggctattg gcttcgctgt gctgctccgg
     2221 gccctgtgga agggcactca ccaagccttt caaccctcca agctaaaggc cattgtagca
     2281 agcaaggcca gccagtttac aggctacgca cagcatgatg cccaagagtt catggctttc
     2341 ttgttggatg ggctgcatga agacctgaat cggatccaaa acaaacccta cacagagact
     2401 gtggactcag atgggcgacc tgacgaggtg gtggctgaag aagcatggca gcggcacaag
     2461 atgaggaacg actcattcat tgtggacctg tttcagggcc agtacaagtc aaagctggtg
     2521 tgccctgtgt gtgccaaggt ctccatcact tttgacccgt tcctttatct gccggtaccc
     2581 ttgccacaaa agcaaaaggt tctccccata tattattttg ccagagagcc ccatagcaaa
     2641 cccatcaagt tcctggtgag cgtcagcaag gagaactcca gcgcaagcga agttttggag
     2701 tccctctctc agagtgtcca cgtgaagcct gagagcctgc gcctggctga ggtaattaag
     2761 aatcgtttcc accgtgtttt cttgccctcc cactcactgg atgctgtgtc gccaacggac
     2821 gtgctcctgt gctttgagct gctatcccca gaattggcta aggagagggt agtggtgcta
     2881 gaggtgcagc agcgccccca ggtacccagc atccctatct ccaagtgtgc agcctgccag
     2941 cggaagcagc aatcagaaga tgaaaaactg aagcgttgta cccgttgcta ccgtgtgggc
     3001 tactgcaacc agttctgtca gaaaacccat tggcctgacc acaaaggcct ctgccgcccc
     3061 gagaacattg gctacccctt cctggtcagt gtacctgctt cacgcctcac ttatgcccgt
     3121 cttgctcagc tactagaagg ttatgcccgg tactctgtga gcgtattcca accacccttc
     3181 cagcctggcc gaatggcttt ggaatcacag agccctggct gtaccacgtt gctttcaacc
     3241 agctctctgg aggctgggga tagtgaaaga gaacccattc agccttctga gctccagttg
     3301 gtgactcctg tggctgaggg ggataccggg gctcaccgca tgtggccacc tgctgatcgg
     3361 ggtcctgtgc ctagcaccag tggaatttct tccgagatgc tggccagtgg gcctatggaa
     3421 gggtgttcct tgcttgctgg tgagagggtg tctcggcctg aagctgctgt gcctgggtac
     3481 cagcactcac gtgagtctgt gagcgcccac acgccccagt tcttcatcta taaaattgat
     3541 gcgtcaagcc gtgagcagcg gctggaggac aaaggggaca ctccactgga gctaggtgat
     3601 gactgtagcc tggctctggt gtggcggaac aacgaacgcc ttcaggagtt tgtgctggtg
     3661 gcctccaagg agctggagtg tgctgaagac ccaggctccg ctggtgaggc tgcccgtgct
     3721 ggccacttca ccctggacca gtgcctcaat ctctttacac ggcctgaagt gctggcacct
     3781 gaggaagcct ggtactgccc acagtgcaaa cagcatcgag aggcctccaa gcagctgctg
     3841 ctgtggcgcc taccaaatgt gctcatcgtg cagctcaagc gtttctcctt tcgtagcttc
     3901 atttggcgtg acaagatcaa tgacttggtg gagtttccag ttcggaacct ggacttgagc
     3961 aagttctgta ttggtcagaa agaggagcag ctgcctagct atgacctgta tgcagtcatc
     4021 aaccactacg gaggcatgat cggcggccac tacactgcct gtgcacggct gcccagtgac
     4081 cgtagtagcc agcgcagtga cgtgggctgg cgcttgtttg atgacagcac ggtgacaaca
     4141 gtagacgaga gccaggtggt gacacgctat gcctatgtgc tcttctaccg tcggcggaac
     4201 tctcctgtgg agaggccccc cagggcagct cacgctgaac accacccaga cctaggccct
     4261 gcagccgagg ctgctgccag ccaggcttcc cggatttggc aggagctcga ggccgaggag
     4321 gagatggtgc ccgaggggcc tgggcctctg ggtccttggg ggccccaaga ctgggtgggg
     4381 cccccgccac gtggccctac cacatcagac gagggctgcc tccgatactt tgtcctgggt
     4441 accgtggcgg ctttggtggc ccttgtgctc aacgtattct atcctctggt atctcagagt
     4501 cgctggagat gagctcccct gcaggcgcct gctgtgagct ggcctaccta tgctcaccag
     4561 gacaggcctg cctttgtggt gggagacagc tctgggctct gagcctcagt gtatgtatct
     4621 ggtgggagac ttgttgggac tacagcctct gtgggggcag agtatcctag ggtgtgtgtc
     4681 catccagttg tctgtcctcc tgtttctctg acccttgtcc tagggcttgg gtaattcgat
     4741 atcaagctaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     4801 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     4861 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     4921 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     4981 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     5041 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     5101 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     5161 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     5221 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     5281 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     5341 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     5401 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     5461 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     5521 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     5581 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     5641 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     5701 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     5761 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     5821 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     5881 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     5941 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     6001 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     6061 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     6121 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     6181 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     6241 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     6301 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     6361 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     6421 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     6481 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     6541 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     6601 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     6661 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     6721 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     6781 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     6841 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     6901 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     6961 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     7021 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     7081 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     7141 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     7201 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     7261 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     7321 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     7381 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     7441 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     7501 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     7561 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     7621 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     7681 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     7741 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     7801 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     7861 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     7921 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     7981 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     8041 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     8101 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     8161 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     8221 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     8281 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     8341 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     8401 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     8461 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     8521 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     8581 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     8641 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     8701 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     8761 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     8821 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     8881 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     8941 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     9001 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     9061 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     9121 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     9181 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     9241 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     9301 aaaa
//
