LOCUS       sequence-311847-        6789 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  for mammalian overexpression of the CS domains of rat USP19.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6789)
  TITLE     rat USP19 constructs
REFERENCE   2  (bases 1 to 6789)
  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-rUSP19CSdomains
FEATURES             Location/Qualifiers
     source          1..6789
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        68..447
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        448..651
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     602..622
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     696..715
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        696..714
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      821..830
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             833..850
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             851..916
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     primer_bind     complement(2211..2228)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2217..2441
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2487..2915
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2574..2593)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2784..2805
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2924..2944)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2929..3258
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3109..3244
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3171..3190
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3325..4119
                     /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(3379..3398)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3989..4008
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4295..4416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4332..4351)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4386..4405
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4465..4481)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4465..4481)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4478..4500)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4489..4505
                     /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(4513..4519,4520..4537,4538..4543))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4558..4579
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4696..4713)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4867..5455)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4947..4966)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5626..6417,6418..6486))
                     /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     6249..6268
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6487..6591)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6666..6685)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
       61 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
      121 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
      181 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
      241 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
      301 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
      361 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
      421 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
      481 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
      541 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
      601 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
      661 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
      721 caagctggct agttaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt
      781 ctgcagatat ccagcacagt ggcggccgct cgagtctaga gccaccatgg cacatcatca
      841 tcatcatcac gactacaaag accatgacgg tgattataaa gatcatgaca tcgattacaa
      901 ggatgacgat gacaagcgcc cgggctgcag gaattcgatg ggtccatgtg ctggcaccag
      961 tgctacaggc ccaaggaggg ggcctccggg attggaggag gcaactagta agaagaaaca
     1021 gaaggatcga gcaaaccagg aaagtaagga tggagatcct aggagagtgt ccatgcctcg
     1081 aaaggagccc accaaagatg aactgttgct cgattggaga cagagttcag ataaggtggt
     1141 tgttaagctg cgcgtgggaa caggtcccat atgtctggag gaagtagatg ctgctttcac
     1201 agacacggac tgtgtggtga ggcttccaga tggtcggcag tggggtggtg tcttctttgc
     1261 tgaaatacaa agttcttgta ccaaagtgca gactcacaag ggtggtcttc tacagttggc
     1321 actaccaaag aaggtgcctc ttctcacgtg gccctctctc ctgaagaaac ctctagggac
     1381 ccaagaactg gtgccaggat tgcggtgcca ggagaacgga caagaactgt ctcccattgc
     1441 cctggagcca ggttctgagc cccgcagagc taaacaggaa gctcgaaacc agaagagggc
     1501 ccagggccgt ggtgaggtag gctcaggggc tagccctggg gcacaggcag ggcccagtgc
     1561 caagagggct gttcatctct gcagagggcc agaaggtgaa gggtccatgg atgggcctgg
     1621 cccccagggt gatgccccat ccttcctgtc cgactcagct acgcaggttg aggctgagga
     1681 gcagctccat gttccaccag tgaaccctca aacaagtctc ttgggctcag agaagaattt
     1741 agcccttttg acagtagaga agacagtgtc ccccaggagt gactcagtct ccccagtcat
     1801 gatccggaac agagaccctg agaaagatga ccatttcaaa gaggagatgg cagttggagc
     1861 tgatcctgca gccttagcgg atgaaccaga gtctatggtg aacctggcat ttgtcaagaa
     1921 tgattcgtat gagaaggggc cggattcagt ggtggtgcac gtgtatgtga aggagagccg
     1981 cagggatacc tctcgagtac ttttccgaga gcaggacttc acgctgatct tccagaccag
     2041 ggatggaaat tttctgaggc tgcatccggg ctgtgggccc cacaccatct tccgctggca
     2101 ggtgaagctc aggaacttga ttgagccaga gcaatgtacg ttttgtttca cggcctctcg
     2161 aatcgatatc tgcctccgga agcggcagag tcagcaaacc cgctgatcag cctcgactgt
     2221 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     2281 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     2341 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     2401 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     2461 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
     2521 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
     2581 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
     2641 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
     2701 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
     2761 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     2821 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     2881 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     2941 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     3001 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     3061 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     3121 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     3181 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     3241 ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa
     3301 gagacaggat gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg
     3361 gccgcttggg tggagaggct attcggctat gactgggcac aacagacaat cggctgctct
     3421 gatgccgccg tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac
     3481 ctgtccggtg ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg
     3541 acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg
     3601 ctattgggcg aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa
     3661 gtatccatca tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca
     3721 ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt
     3781 gtcgatcagg atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc
     3841 aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc
     3901 ttgccgaata tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg
     3961 ggtgtggcgg accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt
     4021 ggcggcgaat gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag
     4081 cgcatcgcct tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga
     4141 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     4201 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     4261 cggggatctc atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg
     4321 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     4381 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc
     4441 tagctagagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     4501 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     4561 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     4621 gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     4681 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     4741 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     4801 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     4861 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     4921 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     4981 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     5041 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     5101 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     5161 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     5221 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     5281 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc
     5341 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     5401 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     5461 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     5521 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     5581 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     5641 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     5701 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     5761 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     5821 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     5881 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     5941 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     6001 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     6061 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     6121 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     6181 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     6241 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     6301 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     6361 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     6421 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     6481 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     6541 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     6601 ccgaaaagtg ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact
     6661 ctcagtacaa tctgctctga tgccgcatag ttaagccagt atctgctccc tgcttgtgtg
     6721 ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta caacaaggca aggcttgacc
     6781 gacaattgc
//
