LOCUS       sequence-193196-        5918 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of CPI-17 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-wt-NLS
FEATURES             Location/Qualifiers
     source          1..5918
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        14..393
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        394..597
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     548..568
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     642..661
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        642..660
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             686..712
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             1190..1210
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     promoter        complement(1250..1268)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1251..1268)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1288..1305)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1294..1518
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1564..1992
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1651..1670)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1861..1882
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2001..2021)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2006..2335
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2186..2321
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2248..2267
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2402..3196
                     /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(2456..2475)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3066..3085
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3370..3491
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3407..3426)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3461..3480
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3540..3556)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3540..3556)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3553..3575)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3564..3580
                     /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(3588..3594,3595..3612,3613..3618))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3633..3654
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3771..3788)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3942..4530)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4022..4041)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4701..5492,5493..5561))
                     /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     5324..5343
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5562..5666)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5741..5760)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agatatacgc gttgacattg attattgact agttattaat agtaatcaat tacggggtca
       61 ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct
      121 ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta
      181 acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta aactgcccac
      241 ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt
      301 aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag
      361 tacatctacg tattagtcat cgctattacc atggtgatgc ggttttggca gtacatcaat
      421 gggcgtggat agcggtttga ctcacgggga tttccaagtc tccaccccat tgacgtcaat
      481 gggagtttgt tttggcacca aaatcaacgg gactttccaa aatgtcgtaa caactccgcc
      541 ccattgacgc aaatgggcgg taggcgtgta cggtgggagg tctatataag cagagctctc
      601 tggctaacta gagaacccac tgcttactgg cttatcgaaa ttaatacgac tcactatagg
      661 gagacccaag cttggtacca tggcgtaccc atacgatgtt ccagattacg ccggatccat
      721 cgatgtcgac gaattcatgg cagctcagcg gctgggcaag cgcgtgctga gcaagctgca
      781 gtctccatcg cgggcccgcg ggccaggggg cagtcccggg gggctgcaga agcggcacgc
      841 gcgcgtcacc gtcaagtatg accggcggga gctgcagcgg cggctggacg tggagaagtg
      901 gatcgacggg cgcctggagg agctgtaccg cggcatggag gcagacatgc ccgatgagat
      961 caacattgat gaattgttgg agttagagag tgaagaggag agaagccgga aaatccaggg
     1021 actcctgaag tcatgtggga aacctgtcga ggacttcatc caggagctgc tggcaaagct
     1081 tcaaggcctc cacaggcagc ccggcctccg ccagccaagc ccctcccacg acggcagcct
     1141 cagccccctc caggaccggg cccggactgc tcaccccggt ggaggtccac ctaaaaagaa
     1201 gcggaaagtg ggtggaggtt gactcgagca tgcatctaga gggccctatt ctatagtgtc
     1261 acctaaatgc tagagctcgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg
     1321 ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt
     1381 cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg
     1441 gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg
     1501 atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc
     1561 cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
     1621 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
     1681 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
     1741 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
     1801 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
     1861 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
     1921 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
     1981 ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc
     2041 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa
     2101 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     2161 catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc
     2221 tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc
     2281 tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct
     2341 cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg
     2401 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     2461 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     2521 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     2581 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     2641 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     2701 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     2761 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     2821 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     2881 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     2941 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     3001 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     3061 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     3121 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     3181 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     3241 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     3301 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     3361 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     3421 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     3481 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     3541 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     3601 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     3661 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     3721 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     3781 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     3841 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     3901 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3961 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     4021 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     4081 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     4141 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     4201 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     4261 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     4321 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     4381 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     4441 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     4501 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     4561 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     4621 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     4681 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     4741 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     4801 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     4861 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     4921 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4981 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     5041 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     5101 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     5161 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     5221 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     5281 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     5341 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     5401 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     5461 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     5521 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     5581 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     5641 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     5701 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     5761 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     5821 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     5881 gggttaggcg ttttgcgctg cttcgcgatg tacgggcc
//
