LOCUS       sequence-198014-        6152 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 6152)
  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 6152)
  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 R133A/P139A (#626)
FEATURES             Location/Qualifiers
     source          1..6152
                     /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             1381..1407
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1471..1489)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1472..1489)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1509..1526)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1515..1739
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1785..2213
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1872..1891)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2082..2103
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2222..2242)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2227..2556
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2407..2542
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2469..2488
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2623..3417
                     /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(2677..2696)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3287..3306
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3591..3712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3628..3647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3682..3701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3761..3777)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3761..3777)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3774..3796)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3785..3801
                     /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(3809..3815,3816..3833,3834..3839))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3854..3875
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3992..4009)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4163..4751)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4243..4262)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4922..5713,5714..5782))
                     /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     5545..5564
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5783..5887)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5962..5981)
                     /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 gggctctcga
     1021 cgccctgcgc gcctgctgat gcctagccag ttggtaagcc aggtgggcaa agaactactg
     1081 gccctggcct acagcgaggc ctgcggcctg cggggggcgc tgctggacgt ctgcgtggag
     1141 cagggcaaga gctgccacag cgtgggccag ctggcactcg accccagcct ggtgcccacc
     1201 ttccagctga ccctcgtgct gcgcctggac tcacgactct ggcccaagat ccaggggctg
     1261 tttagctccg ccaactctcc cttcctccct ggcttcagcc agtccctgac gctgagcact
     1321 ggcttccgag tcatcaagaa gaagctgtac agctcggaac agctgctcat tgaggagtgt
     1381 tacccatacg atgttccaga ttacgcttga gaattctgca gatatccatc acactggcgg
     1441 ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     1501 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1561 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1621 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1681 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     1741 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1801 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1861 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1921 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1981 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     2041 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     2101 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2161 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     2221 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaacagg cagaagtatg
     2281 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2341 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2401 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2461 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     2521 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     2581 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2641 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2701 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2761 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     2821 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2881 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2941 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     3001 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     3061 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3121 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3181 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3241 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3301 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3361 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3421 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3481 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3541 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3601 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3661 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3721 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3781 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3841 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3901 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3961 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     4021 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4081 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4141 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4201 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4261 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4321 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4381 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4441 atcgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4501 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4561 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4621 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4681 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4741 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4801 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4861 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4921 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4981 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     5041 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5101 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5161 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5221 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5281 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5341 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5401 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5461 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5521 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5581 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5641 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5701 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5761 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5821 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5881 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     5941 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     6001 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     6061 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6121 gcttcgcgat gtacgggcca gatatacgcg tt
//
