LOCUS       sequence-335789-        7220 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of 3x-Flag-tagged human PP1 gamma catalytic 
            subunit isoform 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7220)
  TITLE     Balk lab plasmids
REFERENCE   2  (bases 1 to 7220)
  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: Flag-neo-PP1-Gamma
FEATURES             Location/Qualifiers
     source          1..7220
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     126..148
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     140..157
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     141..157
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        318..697
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        698..901
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     852..872
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             931..996
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     polyA_signal    1979..2601
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2178..2197)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(2625..2645)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2643..2940
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             2994..3788
                     /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(3048..3067)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3658..3677
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4440..4574
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4477..4496)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4531..4550
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4579..4598)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4580..4598)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4612..4628)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4612..4628)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4625..4647)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4636..4652
                     /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(4660..4666,4667..4684,4685..4690))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4705..4726
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4843..4860)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5014..5602)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5094..5113)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5773..6564,6565..6633))
                     /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     6396..6415
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6634..6738)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6765..7193
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6852..6871)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7062..7083
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaaccatg gactacaaag accatgacgg tgattataaa
      961 gatcatgaca tcgattacaa ggatgacgat gacaagctta tggcggattt agataaactc
     1021 aacatcgaca gcattatcca acggctgctg gaagtgagag ggtccaagcc tggtaagaat
     1081 gtccagcttc aggagaatga aatcagagga ctgtgcttaa agtctcgtga aatctttctc
     1141 agtcagccta tcctactaga acttgaagca ccactcaaaa tatgtggtga catccatgga
     1201 caatactatg atttgctgcg actttttgag tacggtggtt tcccaccaga aagcaactac
     1261 ctgtttcttg gggactatgt ggacagggga aagcagtcat tggagacgat ctgcctctta
     1321 ctggcctaca aaataaaata tcctgagaat ttttttcttc tcagagggaa ccatgaatgt
     1381 gccagtatca acagaattta tggattttat gatgaatgta aaagaagata caacattaaa
     1441 ctatggaaaa ctttcacaga ctgttttaac tgtttaccga tagcagccat cgtggatgag
     1501 aagatattct gctgtcatgg aggtttatca ccagatcttc aatctatgga gcagattcgg
     1561 cgaattatgc gaccaactga tgtaccagat caaggtcttc tttgtgatct tttgtggtct
     1621 gaccccgata aagatgtctt aggctggggt gaaaatgaca gaggagtgtc cttcacattt
     1681 ggtgcagaag tggttgcaaa atttctccat aagcatgatt tggatcttat atgtagagcc
     1741 catcaggtgg ttgaagatgg atatgaattt tttgcaaaga ggcagttggt cactctgttt
     1801 tctgcgccca attattgcgg agagtttgac aatgcaggtg ccatgatgag tgtggatgaa
     1861 acactaatgt gttcttttca gattttaaag cctgcagaga aaaagaagcc aaatgccacg
     1921 agacctgtaa cgcctccaag gggtatgatc acaaagcaag caaagaaata gggatcccgg
     1981 gtggcatccc tgtgacccct ccccagtgcc tctcctggcc ctggaagttg ccactccagt
     2041 gcccaccagc cttgtcctaa taaaattaag ttgcatcatt ttgtctgact aggtgtcctt
     2101 ctataatatt atggggtgga ggggggtggt atggagcaag gggcaagttg ggaagacaac
     2161 ctgtagggcc tgcggggtct attgggaacc aagctggagt gcagtggcac aatcttggct
     2221 cactgcaatc tccgcctcct gggttcaagc gattctcctg cctcagcctc ccgagttgtt
     2281 gggattccag gcatgcatga ccaggctcag ctaatttttg tttttttggt agagacgggg
     2341 tttcaccata ttggccaggc tggtctccaa ctcctaatct caggtgatct acccaccttg
     2401 gcctcccaaa ttgctgggat tacaggcgtg aaccactgct cccttccctg tccttctgat
     2461 tttaaaataa ctataccagc aggaggacgt ccagacacag cataggctac ctggccatgc
     2521 ccaaccggtg ggacatttga gttgcttgct tggcactgtc ctctcatgcg ttgggtccac
     2581 tcagtagatg cctgttgaat tgggtacgcg gccagcttgg ctgtggaatg tgtgtcagtt
     2641 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     2701 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2761 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2821 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2881 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     2941 aggaattgat cagcttggga tctgatcaag agacaggatg aggatcgttt cgcatgattg
     3001 aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg
     3061 actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg
     3121 ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg
     3181 aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg
     3241 ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc
     3301 tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc
     3361 tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc
     3421 gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc
     3481 aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg
     3541 atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct
     3601 tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt
     3661 tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc
     3721 tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt
     3781 tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc
     3841 acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg
     3901 ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc
     3961 cgggctcgat cccctcgcga gttggttcag ctgctgcctg aggctggacg acctcgcgga
     4021 gttctaccgg cagtgcaaat ccgtcggcat ccaggaaacc agcagcggct atccgcgcat
     4081 ccatgccccc gaactgcagg agtggggagg cacgatggcc gctttggtcg acccggacgg
     4141 gacgctcctg cgcctgatac agaacgaatt gcttgcaggc atctcatgag tgtgtcttcc
     4201 cgttttccgc ctgaggtcac tgcgtggatg gagcgctggc gcctgctgcg cgacggcgag
     4261 ctgctcacca cccactcgcc aagctggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4321 cataggctcc gccgatcata atcagccata ccacatttgt agaggtttta cttgctttaa
     4381 aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt gttgttgtta
     4441 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4501 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4561 atcatgtctg gatcaattcc ctatagtgag tcgtattaaa ttcgtaatca tgtcatagct
     4621 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     4681 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     4741 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     4801 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     4861 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     4921 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     4981 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     5041 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     5101 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     5161 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     5221 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     5281 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     5341 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     5401 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     5461 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     5521 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     5581 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     5641 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     5701 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     5761 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     5821 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     5881 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     5941 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     6001 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     6061 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     6121 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     6181 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     6241 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     6301 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     6361 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     6421 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     6481 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     6541 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     6601 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     6661 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     6721 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgacgcgc cctgtagcgg
     6781 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     6841 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     6901 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     6961 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     7021 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     7081 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     7141 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     7201 aatattaacg cttacaattt
//
