LOCUS       sequence-193197-        5912 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CPI-17 phospho-mimetic cytosolic.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5912)
  TITLE     Gordon Lab plasmids
REFERENCE   2  (bases 1 to 5912)
  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-NES
FEATURES             Location/Qualifiers
     source          1..5912
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        26..405
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        406..609
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     560..580
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     654..673
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        654..672
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             698..724
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1256..1274)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1257..1274)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1294..1311)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1300..1524
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1570..1998
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1657..1676)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1867..1888
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2007..2027)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2012..2341
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2192..2327
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2254..2273
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2408..3202
                     /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(2462..2481)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3072..3091
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3376..3497
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3413..3432)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3467..3486
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3546..3562)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3546..3562)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3559..3581)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3570..3586
                     /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(3594..3600,3601..3618,3619..3624))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3639..3660
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3777..3794)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3948..4536)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4028..4047)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4707..5498,5499..5567))
                     /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     5330..5349
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5568..5672)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5747..5766)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta atagtaatca
       61 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
      121 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
      181 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
      241 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
      301 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
      361 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
      421 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
      481 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
      541 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
      601 agcagagctc tctggctaac tagagaaccc actgcttact ggcttatcga aattaatacg
      661 actcactata gggagaccca agcttggtac catggcgtac ccatacgatg ttccagatta
      721 cgccggatcc atcgatgtcg acgaattcat ggcagctcag cggctgggca agcgcgtgct
      781 gagcaagctg cagtctccat cgcgggcccg cgggccaggg ggcagtcccg gggggctgca
      841 gaagcggcac gcgcgcgtcg aggtcaagta tgaccggcgg gagctgcagc ggcggctgga
      901 cgtggagaag tggatcgacg ggcgcctgga ggagctgtac cgcggcatgg aggcagacat
      961 gcccgatgag atcaacattg atgaattgtt ggagttagag agtgaagagg agagaagccg
     1021 gaaaatccag ggactcctga agtcatgtgg gaaacctgtc gaggacttca tccaggagct
     1081 gctggcaaag cttcaaggcc tccacaggca gcccggcctc cgccagccaa gcccctccca
     1141 cgacggcagc ctcagccccc tccaggaccg ggcccggact gctcaccccc tggccctgaa
     1201 gctggccggc ctggacatcg gcagctgact cgagcatgca tctagagggc cctattctat
     1261 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     1321 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     1381 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     1441 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     1501 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     1561 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     1621 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     1681 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt
     1741 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     1801 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     1861 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     1921 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     1981 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     2041 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     2101 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     2161 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     2221 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     2281 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     2341 aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc
     2401 gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag
     2461 gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg
     2521 gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa
     2581 tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc
     2641 agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc
     2701 ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga
     2761 tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa
     2821 acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct
     2881 ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat
     2941 gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt
     3001 ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta
     3061 tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga
     3121 ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg
     3181 ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg
     3241 cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc
     3301 ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag
     3361 ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc
     3421 atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa
     3481 ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa
     3541 tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata
     3601 cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta
     3661 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3721 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     3781 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     3841 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     3901 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     3961 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     4021 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     4081 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     4141 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     4201 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     4261 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     4321 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     4381 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     4441 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     4501 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     4561 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     4621 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     4681 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     4741 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     4801 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     4861 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     4921 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     4981 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     5041 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     5101 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     5161 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     5221 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     5281 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     5341 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     5401 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     5461 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     5521 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     5581 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     5641 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     5701 gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg
     5761 atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt
     5821 gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc
     5881 tgcttagggt taggcgtttt gcgctgcttc gc
//
