LOCUS       sequence-197987-        5927 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 5927)
  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 5927)
  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 77-232 (#426)
FEATURES             Location/Qualifiers
     source          1..5927
                     /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             1156..1182
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1246..1264)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1247..1264)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1284..1301)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1290..1514
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1560..1988
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1647..1666)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1857..1878
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1997..2017)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2002..2331
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2182..2317
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2244..2263
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2398..3192
                     /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(2452..2471)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3062..3081
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3366..3487
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3403..3422)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3457..3476
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3536..3552)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3536..3552)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3549..3571)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3560..3576
                     /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(3584..3590,3591..3608,3609..3614))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3629..3650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3767..3784)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3938..4526)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4018..4037)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4697..5488,5489..5557))
                     /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     5320..5339
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5558..5662)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5737..5756)
                     /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 caccatgtcg ttgcccgact ttgagctgct cagtgaccct
      721 gaggatgaac acttgtgtgc caacctgatg cagctgctgc aggagagcct ggcccaggcg
      781 cggctgggct ctcgacgccc tgcgcgcctg ctgatgccta gccagttggt aagccaggtg
      841 ggcaaagaac tactgcgcct ggcctacagc gagccgtgcg gcctgcgggg ggcgctgctg
      901 gacgtctgcg tggagcaggg caagagctgc cacagcgtgg gccagctggc actcgacccc
      961 agcctggtgc ccaccttcca gctgaccctc gtgctgcgcc tggactcacg actctggccc
     1021 aagatccagg ggctgtttag ctccgccaac tctcccttcc tccctggctt cagccagtcc
     1081 ctgacgctga gcactggctt ccgagtcatc aagaagaagc tgtacagctc ggaacagctg
     1141 ctcattgagg agtgttaccc atacgatgtt ccagattacg cttgagaatt ctgcagatat
     1201 ccatcacact ggcggccgct cgagcatgca tctagagggc cctattctat agtgtcacct
     1261 aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     1321 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     1381 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     1441 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     1501 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     1561 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     1621 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     1681 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     1741 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     1801 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     1861 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     1921 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     1981 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     2101 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     2161 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     2221 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     2281 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     2341 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     2401 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     2461 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     2521 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     2581 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     2641 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     2701 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     2761 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     2821 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     2881 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     2941 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3001 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3061 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     3121 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     3181 gagttcttct gagcgggact ctggggttcg aaatgaccga ccaagcgacg cccaacctgc
     3241 catcacgaga tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt
     3301 tccgggacgc cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc
     3361 accccaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     3421 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     3481 tatcttatca tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat
     3541 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     3601 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     3661 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     3721 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     3781 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3841 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3901 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3961 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4021 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4081 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4141 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4201 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4261 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     4321 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     4381 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4441 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4501 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4561 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     4621 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     4681 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     4741 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     4801 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     4861 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     4921 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4981 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5041 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5101 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5161 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5221 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     5281 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     5341 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     5401 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     5461 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     5521 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     5581 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     5641 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gtcgacggat
     5701 cgggagatct cccgatcccc tatggtgcac tctcagtaca atctgctctg atgccgcata
     5761 gttaagccag tatctgctcc ctgcttgtgt gttggaggtc gctgagtagt gcgcgagcaa
     5821 aatttaagct acaacaaggc aaggcttgac cgacaattgc atgaagaatc tgcttagggt
     5881 taggcgtttt gcgctgcttc gcgatgtacg ggccagatat acgcgtt
//
