LOCUS       sequence-198002-        6125 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 6125)
  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 6125)
  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 N179-193 (#437)
FEATURES             Location/Qualifiers
     source          1..6125
                     /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             1354..1380
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1444..1462)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1445..1462)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1482..1499)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1488..1712
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1758..2186
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1845..1864)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2055..2076
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2195..2215)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2200..2529
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2380..2515
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2442..2461
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2596..3390
                     /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(2650..2669)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3260..3279
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3564..3685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3601..3620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3655..3674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3734..3750)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3734..3750)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3747..3769)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3758..3774
                     /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(3782..3788,3789..3806,3807..3812))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3827..3848
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3965..3982)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4136..4724)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4216..4235)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4895..5686,5687..5755))
                     /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     5518..5537
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5756..5860)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5935..5954)
                     /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 cgcctggcct acagcgagcc gtgcggcctg cggggggcgc tgctggacgt ctgcgtggag
     1141 cagggcaaga gctgccacag cgtgggccag ctggcactcg accccagcct ggtgcccacc
     1201 ttccagctga ccctcgtgaa tgctgctata cgatcgagct ccgccaactc tcccttcctc
     1261 cctggcttca gccagtccct gacgctgagc actggcttcc gagtcatcaa gaagaagctg
     1321 tacagctcgg aacagctgct cattgaggag tgttacccat acgatgttcc agattacgct
     1381 tgagaattct gcagatatcc atcacactgg cggccgctcg agcatgcatc tagagggccc
     1441 tattctatag tgtcacctaa atgctagagc tcgctgatca gcctcgactg tgccttctag
     1501 ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac
     1561 tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca
     1621 ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag
     1681 caggcatgct ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg
     1741 ctctaggggg tatccccacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1801 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1861 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1921 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1981 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     2041 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggc
     2101 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     2161 gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga
     2221 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     2281 accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc
     2341 aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc
     2401 agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag
     2461 gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc
     2521 ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca agagacagga
     2581 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     2641 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     2701 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     2761 gccctgaatg aactgcagga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     2821 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     2881 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     2941 atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac
     3001 caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag
     3061 gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag
     3121 gcgcgcatgc ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat
     3181 atcatggtgg aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg
     3241 gaccgctatc aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa
     3301 tgggctgacc gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc
     3361 ttctatcgcc ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc
     3421 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3481 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3541 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3601 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3661 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     3721 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3781 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3841 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3901 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3961 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     4021 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     4081 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     4141 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     4201 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     4261 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     4321 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     4381 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     4441 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4501 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4561 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4621 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4681 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4741 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4801 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4861 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4921 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4981 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     5041 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     5101 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     5161 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     5221 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     5281 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     5341 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     5401 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     5461 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5521 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5581 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5641 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5701 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5761 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5821 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5881 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     5941 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     6001 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     6061 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     6121 gcgtt
//
