LOCUS       sequence-211672-        7391 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  URT-type vector for Glu-mI-kappa-B-alpha expression in Mammalian 
            cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7391)
  AUTHORS   Piatkov KI, Oh JH, Liu Y, Varshavsky A
  TITLE     Calpain-generated natural protein fragments as short-lived 
            substrates of the N-end rule pathway.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Mar 4;111(9):E817-26. doi: 
            10.1073/pnas.1401639111. Epub 2014 Feb 18.
  PUBMED    24550490
REFERENCE   2  (bases 1 to 7391)
  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"; doi: 
            "10.1073/pnas.1401639111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-03-4- 4"; volume: "111"; issue: "9"; pages: "E817-26"
COMMENT     Sequence Label: pJO427
FEATURES             Location/Qualifiers
     source          1..7391
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             664..687
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1000..1560
                     /codon_start=1
                     /gene="Mus musculus Dhfr"
                     /product="mouse dihydrofolate reductase"
                     /label=DHFR
                     /translation="MVRPLNCIVAVSQNMGIGKNGDLPWPPLRNEFKYFQRMTTTSSVE
                     GKQNLVIMGRKTWFSIPEKNRPLKDRINIVLSRELKEPPRGAHFLAKSLDDALRLIEQP
                     ELASKVDMVWIVGGSSVYQEAMNQPGHLRLFVTRIMQEFESDTFFPEIDLGKYKLLPEY
                     PGVLSEVQEEKGIKYKFEVYEKKD"
     CDS             1567..1593
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2665..2688
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2710..2728)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2711..2728)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2748..2765)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2754..2978
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3024..3452
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3111..3130)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3321..3342
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3461..3481)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3466..3795
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3646..3781
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3708..3727
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3862..4656
                     /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(3916..3935)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4526..4545
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4830..4951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4867..4886)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4921..4940
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5000..5016)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5000..5016)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5013..5035)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5024..5040
                     /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(5048..5054,5055..5072,5073..5078))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5093..5114
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5231..5248)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5402..5990)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5482..5501)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6161..6952,6953..7021))
                     /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     6784..6803
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7022..7126)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7201..7220)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 atggactaca aggacgacga tgacaagggt gcacgacagg tttcccgact taatcgcctt
      721 gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct
      781 tcccaacagt tgcgcagcct gaatggcgaa tggcgctttg cctggtttcc ggcaccagaa
      841 gcggtgccgg aaagctggct ggagtgcgat cttcctgagg ccgatactgt cgtcgtcccc
      901 tcaaactggc agatgcacgg ttacgatgcg cccatctaca ccaacgtgac ctatcccatt
      961 acggtcaatc cgccgtttgt tcccacggag aatggatcca tggttcgacc attgaactgc
     1021 atcgtcgccg tgtcccaaaa tatggggatt ggcaagaacg gagacctacc ctggcctccg
     1081 ctcaggaacg agttcaagta cttccaaaga atgaccacaa cctcttcagt ggaaggtaaa
     1141 cagaatctgg tgattatggg taggaaaacc tggttctcca ttcctgagaa gaatcgacct
     1201 ttaaaggaca gaattaatat agttctcagt agagaactca aagaaccacc acgaggagct
     1261 cattttcttg ccaaaagttt ggatgatgcc ttaagactta ttgaacaacc ggaattggca
     1321 agtaaagtag acatggtttg gatagtcgga ggcagttctg tttaccagga agccatgaat
     1381 caaccaggcc acctcagact ctttgtgaca aggatcatgc aggaatttga aagtgacacg
     1441 tttttcccag aaattgattt ggggaaatat aaacttctcc cagaataccc aggcgtcctc
     1501 tctgaggtcc aggaggaaaa aggcatcaag tataagtttg aagtctacga gaagaaagac
     1561 ggtacctacc catacgatgt cccagattac gccgctttcc ttgcgggtag tgggtcgacc
     1621 atgcagattt tcgtcaagac tttgaccggt aaaaccataa cattggaagt tgaatcttcc
     1681 gataccatcg acaacgttaa gtcgaaaatt caagacaagg aaggtatccc tccagatcaa
     1741 caaagattga tctttgccgg taggcagcta gaagacggta gaacgctgtc tgattacaac
     1801 attcagaagg agtccacctt acatcttgtg ctaaggctcc gcggaggaga gatccgcctg
     1861 cagccgcagg aggcgccgct ggccgccgag ccctggaagc agcagctcac ggaggacgga
     1921 gactcgttcc tgcacttggc aatcatccac gaagagaagc cgctgaccat ggaagtcatt
     1981 ggtcaggtga agggagacct ggccttcctc aacttccaga acaacctgca gcagactcca
     2041 ctccacttgg ctgtgatcac caaccagcca ggaattgctg aggcacttct gaaagctggc
     2101 tgtgatcctg agctccgaga ctttcgagga aatacccctc tacatcttgc ctgtgagcag
     2161 ggctgcctgg ccagtgtagc agtcttgacg cagacctgca caccccagca tctccactcc
     2221 gtcctgcagg ccaccaacta caatggccac acgtgtctgc acctagcctc tatccacggc
     2281 tacctggcca tcgtggagca cttggtgact ttgggtgctg atgtcaacgc tcaggagccc
     2341 tgcaatggcc ggacagccct ccaccttgcg gtggacctgc agaatcctga cctggtttcg
     2401 ctcttgttga aatgtggggc tgatgtcaac agggtaacct accaaggcta ctccccctac
     2461 cagcttacct ggggccgccc aagtacccgg atacagcagc agctgggcca gctgaccctg
     2521 gaaaatctcc agatgctacc cgagagcgag gatgaggaga gctatgacac ggagtcagaa
     2581 ttcacagagg atgagctgcc ctatgatgac tgtgtgtttg gaggccagcg tctgacatta
     2641 atggctatgt ccatgatcga tatcgactac aaagacgatg acgacaaagg ttaatctaga
     2701 gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct cgactgtgcc
     2761 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     2821 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag
     2881 gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga
     2941 caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag
     3001 ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3061 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3121 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     3181 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     3241 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     3301 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3361 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3421 tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg
     3481 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3541 tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3601 catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact
     3661 ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag
     3721 gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc
     3781 ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag
     3841 acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc
     3901 gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat
     3961 gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg
     4021 tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg
     4081 ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta
     4141 ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta
     4201 tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc
     4261 gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc
     4321 gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg
     4381 ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg
     4441 ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt
     4501 gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc
     4561 ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc
     4621 atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga
     4681 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg
     4741 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     4801 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     4861 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4921 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4981 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     5041 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     5101 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     5161 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     5221 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5281 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5341 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5401 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5461 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5521 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5581 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5641 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5701 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5761 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5821 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5881 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5941 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     6001 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     6061 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6121 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6181 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6241 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6301 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6361 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6421 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6481 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6541 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6601 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6661 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6721 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6781 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6841 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6901 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6961 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     7021 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     7081 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7141 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag
     7201 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga
     7261 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa
     7321 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggccag
     7381 atatacgcgt t
//
