LOCUS       sequence-134170-        5536 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5536)
  AUTHORS   Vander Haar E, Lee SI, Bandhakavi S, Griffin TJ, Kim DH
  TITLE     Insulin signalling to mTOR mediated by the Akt/PKB substrate PRAS40.
  JOURNAL   Nat Cell Biol. 2007 Mar . 9(3):316-23.
  PUBMED    17277771
REFERENCE   2  (bases 1 to 5536)
  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"; journalName: "Nat Cell 
            Biol. 2007 Mar . 9(3):316-23."
COMMENT     Sequence Label: pRK5-HA-PRAS40
FEATURES             Location/Qualifiers
     source          1..5536
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     enhancer        250..629
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        630..833
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     784..804
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     830..854
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        1064..1082
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1064..1081
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     1095..1114
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     regulatory      1199..1208
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1211..1237
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     regulatory      2072..2081
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     polyA_signal    2085..2219
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2122..2141)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2176..2195
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     promoter        2288..2645
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2288..2479
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2311..2331)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2496..2631
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(2665..2682)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2665..2681)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2674..2696)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2894..3349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2981..3000)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3191..3212
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3366..3385)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3485..3507
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3545..3563)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3631..3735
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3736..3804,3805..4596)
                     /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     complement(3954..3973)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4767..5355
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5256..5275
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5509..5526
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga catgattacg aattaattcg
      241 agctcgcccg acattgatta ttgactagtt attaatagta atcaattacg gggtcattag
      301 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
      361 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      421 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      481 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      541 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      601 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      661 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      721 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      781 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
      841 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
      901 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
      961 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     1021 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     1081 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     1141 cggttctatc gattgaattc ccgaaccgac agtcggtctc ttcaccaagg ccattcgcgc
     1201 caccatgggc tacccctatg atgtgcctga ctacgcagat ctcaatggtg gtggtggtgg
     1261 gtcgacgatg gcgtcgggtc gccctgagga gctgtgggag gccgtggtgg gggccgctga
     1321 gcgcttccgg gcccggactg gcacggagct ggtgctgctg accgcggccc cgccgccacc
     1381 accccgcccg ggcccctgtg cctatcctgc ccatggtcga ggagccctgg cggaggcagc
     1441 gcgccgttgc ctccacgaca tcgcactggc ccacagggct gccactgctg ctcggcctcc
     1501 tgcgccccca ccagcaccac agccacccag tcccacaccc agcccacccc ggcctaccct
     1561 ggccagagag gacaacgagg aggacgagga tgagcccaca gagacagaga cctccgggga
     1621 gcagctgggc attagtgata atggagggct ctttgtgatg gatgaggacg ccaccctcca
     1681 ggaccttccc cccttctgtg agtcagaccc cgagagtaca gatgatggca gcctgagcga
     1741 ggagaccccc gccggccccc ccacctgctc agtgccccca gcctcagccc tacccacaca
     1801 gcagtacgcc aagtccctgc ctgtgtctgt gcccgtctgg ggcttcaagg agaagaggac
     1861 agaggcgcgg tcatcagatg aggagaatgg gccgccctct tcgcccgacc tggaccgcat
     1921 cgcggcgagc atgcgcgcgc tggtgctgcg agaggccgag gacacccagg tcttcgggga
     1981 cctgccacgg ccgcggctta acaccagcga cttccagaag ctgaagcgga aatattgagc
     2041 ggccgctaag taagtaagga tccccagctt ggccgccatg gcccaacttg tttattgcag
     2101 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     2161 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcg
     2221 ggaattaatt cggcgcagca ccatggcctg aaataacctc tgaaagagga acttggttag
     2281 gtaccttctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     2341 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     2401 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     2461 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     2521 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     2581 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     2641 gcaaaaagct gttaacagct tggcactggc cgtcgtttta caacgtcgtg actgggaaaa
     2701 ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa
     2761 tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg
     2821 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcatacgtca
     2881 aagcaaccat agtacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     2941 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3001 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3061 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt gggtgatggt
     3121 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3181 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcgggctat
     3241 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3301 taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttt atggtgcact
     3361 ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc gccaacaccc
     3421 gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca agctgtgacc
     3481 gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg cgcgagacga
     3541 aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat ggtttcttag
     3601 acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa
     3661 atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat
     3721 tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg
     3781 gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa
     3841 gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt
     3901 gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt
     3961 ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat
     4021 tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg
     4081 acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta
     4141 cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat
     4201 catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag
     4261 cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa
     4321 ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca
     4381 ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc
     4441 ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt
     4501 atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc
     4561 gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat
     4621 atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt
     4681 tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac
     4741 cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc
     4801 ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca
     4861 actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta
     4921 gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct
     4981 ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg
     5041 gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc
     5101 acacagccca gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta
     5161 tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg
     5221 gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt
     5281 cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg
     5341 cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg
     5401 ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc
     5461 gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg
     5521 agcgaggaag cggaag
//
