LOCUS       sequence-423105-        6749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Vector for transient expression of PPP2CB gene with C-terminal HA 
            tag, gene flanked by att sites for gateway cloning.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6749)
  TITLE     Chromosome 8p-engineering reveals increased metastasis targetable by
            patient-specific synthetic lethality
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6749)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pDEST26-PPP2CB-C-HA
FEATURES             Location/Qualifiers
     source          1..6749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        11..314
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        315..518
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     469..489
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     515..539
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     protein_bind    636..660
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     regulatory      1157..1166
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             complement(1513..1524)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     protein_bind    complement(1588..1612)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1617..1643
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(1674..1693)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1675..1693)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1704..1721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1704..1720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1713..1735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     polyA_signal    2125..2259
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2162..2181)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2216..2235
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      2385..2840
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2472..2491)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2682..2703
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2957..3314
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2957..3148
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2980..3000)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3165..3300
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3365..4159
                     /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(3419..3438)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4029..4048
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4223..4271
                     /note="synthetic polyadenylation signal"
     primer_bind     complement(4312..4331)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4431..4453
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     promoter        4577..4681
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4682..4750,4751..5542)
                     /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     complement(4900..4919)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5713..6301
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6202..6221
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6455..6472
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        complement(6546..6650)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 agatgcatgt cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
       61 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      121 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      181 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      241 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      301 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      361 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      421 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      481 gtaggcgtgt acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatcg
      541 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      601 tccggactct agcctaggcc gcggagctcg agtcaacaag tttgtacaaa aaagttggca
      661 tggacgacaa ggcgttcacc aaggagctgg accagtgggt cgagcagctg aacgagtgta
      721 agcagctgaa cgagaaccaa gtgcggacgc tgtgcgagaa ggcaaaggaa attttaacaa
      781 aagaatcaaa tgtgcaagag gttcgttgcc ctgttactgt ctgtggagat gtgcatggtc
      841 aatttcatga tcttatggaa ctctttagaa ttggtggaaa atcaccggat acaaactact
      901 tattcatggg tgactatgta gacagaggat attattcagt ggagactgtg actcttcttg
      961 tagcattaaa ggtgcgttat ccagaacgca ttacaatatt gagaggaaat cacgaaagcc
     1021 gacaaattac ccaagtatat ggcttttatg atgaatgtct gcgaaagtat gggaatgcca
     1081 acgtttggaa atattttaca gatctctttg attatcttcc acttacagct ttagtagatg
     1141 gacagatatt ctgcctccat ggtggcctct ctccatccat agacacactg gatcatataa
     1201 gagccctgga tcgtttacag gaagttccac atgagggccc aatgtgtgat ctgttatggt
     1261 cagatccaga tgatcgtggt ggatggggta tttcaccacg tggtgctggc tacacatttg
     1321 gacaagacat ttctgaaacc tttaaccatg ccaatggtct cacactggtt tctcgtgccc
     1381 accagcttgt aatggaggga tacaattggt gtcatgatcg gaatgtggtt accattttca
     1441 gtgcacccaa ttactgttat cgttgtggga accaggctgc tatcatggaa ttagatgaca
     1501 ctttaaaata ttccttcctt caatttgacc cagcgcctcg tcgtggtgag cctcatgtta
     1561 cacggcgcac cccagactac ttcctatacc caactttctt gtacaaagtg gttgattacc
     1621 catacgatgt tccagattac gcttgacgcg tgcatgcgac gtcatagctc tctccctata
     1681 gtgagtcgta ttataagcta ggcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     1741 tgctagcttg ggatctttgt gaaggaacct tacttctgtg gtgtgacata attggacaaa
     1801 ctacctacag agatttaaag ctctaaggta aatataaaat ttttaagtgt ataatgtgtt
     1861 aaactagctg catatgcttg ctgcttgaga gttttgctta ctgagtatga tttatgaaaa
     1921 tattatacac aggagctagt gattctaatt gtttgtgtat tttagattca cagtcccaag
     1981 gctcatttca ggcccctcag tcctcacagt ctgttcatga tcataatcag ccataccaca
     2041 tttgtagagg ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat
     2101 aaaatgaatg caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa
     2161 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2221 ttgtccaaac tcatcaatgt atcttatcat gtctggatcg atcctgcatt aatgaatcgg
     2281 ccaacgcgcg gggagaggcg gtttgcgtat tggctggcgt aatagcgaag aggcccgcac
     2341 cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg
     2401 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     2461 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     2521 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     2581 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2641 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2701 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2761 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     2821 aatattaacg cttacaattt cctgatgcgg tattttctcc ttacgcatct gtgcggtatt
     2881 tcacaccgca tacgcggatc tgcgcagcac catggcctga aataacctct gaaagaggaa
     2941 cttggttagg taccttctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt
     3001 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3061 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     3121 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     3181 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     3241 ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     3301 taggcttttg caaaaagctt gattcttctg acacaacagt ctcgaactta aggctagagc
     3361 caccatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3421 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3481 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     3541 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     3601 tgtgctcgac gttgtcactg aagcgggaag ggattggctg ctattgggcg aagtgccggg
     3661 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     3721 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     3781 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     3841 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     3901 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     3961 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4021 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4081 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4141 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4201 aacctgccat cacgatggcc gcaataaaat atctttattt tcattacatc tgtgtgttgg
     4261 ttttttgtgt gaatcgatag cgataaggat ccgcgtatgg tgcactctca gtacaatctg
     4321 ctctgatgcc gcatagttaa gccagccccg acacccgcca acacccgctg acgcgccctg
     4381 acgggcttgt ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg
     4441 catgtgtcag aggttttcac cgtcatcacc gaaacgcgcg agacgaaagg gcctcgtgat
     4501 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     4561 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     4621 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     4681 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     4741 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     4801 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     4861 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     4921 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     4981 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5041 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5101 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5161 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5221 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5281 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5341 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     5401 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5461 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     5521 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     5581 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     5641 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     5701 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     5761 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     5821 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     5881 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     5941 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6001 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6061 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     6121 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     6181 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     6241 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     6301 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     6361 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     6421 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     6481 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagagc
     6541 ttgcaattcg cgctttttca atattattga agcatttatc agggttattg tctcatgagc
     6601 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     6661 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     6721 aggcgtatta cgaggccctt tcacgcatt
//
