LOCUS       sequence-193198-        5918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CPI-17 phospho-mimetic nuclear.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5918)
  TITLE     Gordon Lab plasmids
REFERENCE   2  (bases 1 to 5918)
  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: HA-CPI-17-T38E-NLS
FEATURES             Location/Qualifiers
     source          1..5918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        11..390
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        391..594
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     545..565
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     639..658
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        639..657
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             683..709
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1187..1207
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1247..1265)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1248..1265)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1285..1302)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1291..1515
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1561..1989
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1648..1667)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1858..1879
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1998..2018)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2003..2332
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2183..2318
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2245..2264
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2399..3193
                     /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(2453..2472)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3063..3082
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3367..3488
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3404..3423)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3458..3477
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3537..3553)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3537..3553)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3550..3572)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3561..3577
                     /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(3585..3591,3592..3609,3610..3615))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3630..3651
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3768..3785)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3939..4527)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4019..4038)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4698..5489,5490..5558))
                     /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     5321..5340
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5559..5663)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5738..5757)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
       61 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
      121 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
      181 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
      241 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
      301 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
      361 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
      421 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
      481 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
      541 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
      601 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
      661 acccaagctt ggtaccatgg cgtacccata cgatgttcca gattacgccg gatccatcga
      721 tgtcgacgaa ttcatggcag ctcagcggct gggcaagcgc gtgctgagca agctgcagtc
      781 tccatcgcgg gcccgcgggc cagggggcag tcccgggggg ctgcagaagc ggcacgcgcg
      841 cgtcgaggtc aagtatgacc ggcgggagct gcagcggcgg ctggacgtgg agaagtggat
      901 cgacgggcgc ctggaggagc tgtaccgcgg catggaggca gacatgcccg atgagatcaa
      961 cattgatgaa ttgttggagt tagagagtga agaggagaga agccggaaaa tccagggact
     1021 cctgaagtca tgtgggaaac ctgtcgagga cttcatccag gagctgctgg caaagcttca
     1081 aggcctccac aggcagcccg gcctccgcca gccaagcccc tcccacgacg gcagcctcag
     1141 ccccctccag gaccgggccc ggactgctca ccccggtgga ggtccaccta aaaagaagcg
     1201 gaaagtgggt ggaggttgac tcgagcatgc atctagaggg ccctattcta tagtgtcacc
     1261 taaatgctag agctcgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     1321 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     1381 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     1441 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     1501 cggtgggctc tatggcttct gaggcggaaa gaaccagctg gggctctagg gggtatcccc
     1561 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     1621 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     1681 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     1741 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     1801 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     1861 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     1921 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     1981 acgcgaatta attctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2041 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2101 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2161 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2221 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct ctgcctctga
     2281 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctccc
     2341 gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat cgtttcgcat
     2401 gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga ggctattcgg
     2461 ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc
     2521 gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga atgaactgca
     2581 ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg cagctgtgct
     2641 cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc cggggcagga
     2701 tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg atgcaatgcg
     2761 gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga aacatcgcat
     2821 cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc tggacgaaga
     2881 gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg
     2941 cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg
     3001 ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct atcaggacat
     3061 agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg accgcttcct
     3121 cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc gccttcttga
     3181 cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac gcccaacctg
     3241 ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     3301 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     3361 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3421 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     3481 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     3541 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     3601 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     3661 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     3721 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     3781 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3841 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3901 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3961 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4021 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4081 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4141 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4201 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4261 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4321 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     4381 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4441 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4501 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4561 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     4621 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     4681 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     4741 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     4801 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     4861 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     4921 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     4981 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     5041 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     5101 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     5161 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     5221 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     5281 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     5341 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     5401 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     5461 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     5521 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     5581 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     5641 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga
     5701 tcgggagatc tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat
     5761 agttaagcca gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca
     5821 aaatttaagc tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg
     5881 ttaggcgttt tgcgctgctt cgcgatgtac gggccaga
//
