LOCUS       sequence-197990-        6032 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 6032)
  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 6032)
  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 1-192 (#421)
FEATURES             Location/Qualifiers
     source          1..6032
                     /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             1261..1287
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     promoter        complement(1351..1369)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1352..1369)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1389..1406)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1395..1619
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1665..2093
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1752..1771)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1962..1983
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2102..2122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2107..2436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2287..2422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2349..2368
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2503..3297
                     /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(2557..2576)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3167..3186
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3471..3592
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3508..3527)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3562..3581
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3641..3657)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3641..3657)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3654..3676)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3665..3681
                     /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(3689..3695,3696..3713,3714..3719))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3734..3755
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3872..3889)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4043..4631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4123..4142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4802..5593,5594..5662))
                     /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     5425..5444
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5663..5767)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5842..5861)
                     /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 ccctcgtgct gcgcctggac tcacgactct ggcccaagat ccaggggctg
     1261 tacccatacg atgttccaga ttacgcttga gaattctgca gatatccatc acactggcgg
     1321 ccgctcgagc atgcatctag agggccctat tctatagtgt cacctaaatg ctagagctcg
     1381 ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc cctcccccgt
     1441 gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa atgaggaaat
     1501 tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg ggcaggacag
     1561 caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg gctctatggc
     1621 ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc cctgtagcgg
     1681 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     1741 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     1801 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     1861 cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac
     1921 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     1981 tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat
     2041 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attaattctg
     2101 tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg cagaagtatg
     2161 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     2221 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     2281 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     2341 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     2401 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     2461 attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     2521 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     2581 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     2641 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg
     2701 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     2761 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     2821 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     2881 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     2941 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     3001 ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg
     3061 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     3121 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     3181 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     3241 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     3301 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3361 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3421 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3481 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3541 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3601 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3661 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3721 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3781 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3841 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3901 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     3961 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4021 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4081 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4141 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4201 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4261 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4321 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4381 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4441 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4501 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4561 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4621 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4681 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4741 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4801 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4861 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4921 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     4981 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5041 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5101 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5161 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5221 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5281 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5341 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5401 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5461 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5521 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5581 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5641 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5701 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5761 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     5821 tcccctatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     5881 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     5941 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
     6001 gcttcgcgat gtacgggcca gatatacgcg tt
//
