LOCUS       sequence-197994-        6077 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  mammalian expression.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6077)
  AUTHORS   Vega-Rubin-de-Celis S, Abdallah Z, Kinch L, Grishin NV, Brugarolas 
            J, Zhang X
  TITLE     Structural analysis and functional implications of the negative 
            mTORC1 regulator REDD1.
  JOURNAL   Biochemistry. 2010 Mar 23;49(11):2491-501. doi: 10.1021/bi902135e.
  PUBMED    20166753
REFERENCE   2  (bases 1 to 6077)
  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"; journalName: 
            "Biochemistry"; date: "2010-03-23- 23"; volume: "49"; issue: "11"; 
            pages: "2491-501"
COMMENT     Sequence Label: pcDNA3-HA-REDD1 111-135 (#190)
FEATURES             Location/Qualifiers
     source          1..6077
                     /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             1306..1332
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1396..1414)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1397..1414)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1434..1451)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1440..1664
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1710..2138
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1797..1816)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2007..2028
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2147..2167)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2152..2481
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2332..2467
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2394..2413
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2548..3342
                     /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(2602..2621)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3212..3231
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3516..3637
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3553..3572)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3607..3626
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3686..3702)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3686..3702)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3699..3721)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3710..3726
                     /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(3734..3740,3741..3758,3759..3764))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3779..3800
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3917..3934)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4088..4676)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4168..4187)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4847..5638,5639..5707))
                     /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     5470..5489
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5708..5812)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5887..5906)
                     /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 ggtaccgagc tcggatccgc caccatgcct agcctttggg accgcttctc gtcgtcgtcc
      721 acctcctctt cgccctcgtc cttgccccga actcccaccc cagatcggcc gccgcgctca
      781 gcctgggggt cggcgacccg ggaggagggg tttgaccgct ccacgagcct ggagagctcg
      841 gactgcgagt ccctggacag cagcaacagt ggcttcgggc cggaggaaga cacggcttac
      901 ctggatgggg tgtcgttgcc cgactttgag ctgctcagtg accctgagga tgaacacttg
      961 tgtgccaacc tgatgcagct gctgcaggag agcctggccc aggcgcggct gggctacagc
     1021 gagccgtgcg gcctgcgggg ggcgctgctg gacgtctgcg tggagcaggg caagagctgc
     1081 cacagcgtgg gccagctggc actcgacccc agcctggtgc ccaccttcca gctgaccctc
     1141 gtgctgcgcc tggactcacg actctggccc aagatccagg ggctgtttag ctccgccaac
     1201 tctcccttcc tccctggctt cagccagtcc ctgacgctga gcactggctt ccgagtcatc
     1261 aagaagaagc tgtacagctc ggaacagctg ctcattgagg agtgttaccc atacgatgtt
     1321 ccagattacg cttgagaatt ctgcagatat ccatcacact ggcggccgct cgagcatgca
     1381 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     1441 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     1501 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     1561 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     1621 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     1681 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     1741 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     1801 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     1861 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     1921 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     1981 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2041 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     2101 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2161 ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2221 aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa
     2281 agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc
     2341 ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat
     2401 gcagaggccg aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt
     2461 ggaggcctag gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat
     2521 caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct
     2581 ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc
     2641 tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc
     2701 gacctgtccg gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc
     2761 acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg
     2821 ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag
     2881 aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc
     2941 ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt
     3001 cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc
     3061 gccaggctca aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc
     3121 tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg
     3181 ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag
     3241 cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg
     3301 cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg
     3361 aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct
     3421 tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc
     3481 gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg
     3541 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3601 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct
     3661 ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc
     3721 tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat
     3781 gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc
     3841 tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg
     3901 ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     3961 cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag
     4021 gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc
     4081 tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc
     4141 agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc
     4201 tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt
     4261 cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg
     4321 ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat
     4381 ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag
     4441 ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt
     4501 ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc
     4561 cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta
     4621 gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag
     4681 atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga
     4741 ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa
     4801 gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa
     4861 tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc
     4921 ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga
     4981 taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa
     5041 gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt
     5101 gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg
     5161 ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc
     5221 aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg
     5281 gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag
     5341 cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt
     5401 actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt
     5461 caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac
     5521 gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac
     5581 ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag
     5641 caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa
     5701 tactcatact cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga
     5761 gcggatacat atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc
     5821 cccgaaaagt gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac
     5881 tctcagtaca atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt
     5941 gttggaggtc gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac
     6001 cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg
     6061 ggccagatat acgcgtt
//
