LOCUS       sequence-248176-        7294 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 7294)
  AUTHORS   Nemoto S, Finkel T
  TITLE     Redox regulation of forkhead proteins through a p66shc-dependent 
            signaling pathway.
  JOURNAL   Science. 2002 Mar 29. 295(5564):2450-2.
  PUBMED    11884717
REFERENCE   2  (bases 1 to 7294)
  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: "Science. 
            2002 Mar 29. 295(5564):2450-2."
COMMENT     Sequence Label: pcDNA3.1his p66shc (corrected)
FEATURES             Location/Qualifiers
     source          1..7294
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        88..467
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        468..671
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     622..642
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     716..735
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        716..734
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             785..802
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             806..838
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(842..850,851..865)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(2738..2755)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2744..2968
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3014..3442
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3101..3120)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3311..3332
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3451..3471)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3456..3785
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3636..3771
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3698..3717
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3852..4646
                     /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(3906..3925)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4516..4535
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4820..4941
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4857..4876)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4911..4930
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4990..5006)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4990..5006)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5003..5025)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5014..5030
                     /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(5038..5044,5045..5062,5063..5068))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5083..5104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5221..5238)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5392..5980)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5472..5491)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6151..6942,6943..7011))
                     /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     6774..6793
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7012..7116)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7191..7210)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aaggcttgac cgacaattgc atgaagaatc tgcttagggt taggcgtttt gcgctgcttc
       61 gcgatgtacg ggccagatat acgcgttgac attgattatt gactagttat taatagtaat
      121 caattacggg gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg
      181 taaatggccc gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt
      241 atgttcccat agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac
      301 ggtaaactgc ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg
      361 acgtcaatga cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact
      421 ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt
      481 ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc
      541 ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc
      601 gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata
      661 taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc gaaattaata
      721 cgactcacta tagggagacc caagctggct agcgtttaaa cttaagctta ccatgggggg
      781 ttctcatcat catcatcatc atggtatggc tagcatgact ggtggacagc aaatgggtcg
      841 ggatctgtac gacgatgacg ataaggtacc gagctcggat ccactagtcc agtgtggtgg
      901 aattcggctt atggatctcc tgccccccaa gcccaagtac aatccactcc ggaatgagtc
      961 tctgtcatcg ctggaggaag gggcttctgg gtccaccccc ccggaggagc tgccttcccc
     1021 atcagcttca tccctggggc ccatcctgcc tcctctgcct ggggacgata gtcccactac
     1081 cctgtgctcc ttcttccccc ggatgagcaa cctgaggctg gccaacccgg ctggggggcg
     1141 cccagggtct aagggggagc caggaagggc agctgatgat ggggagggga tcgtaggggc
     1201 agccatgcca gactcaggcc ccctacccct cctccaggac atgaacaagc tgagtggagg
     1261 cggcgggcgc aggactcggg tggaaggggg ccagcttggg ggcgaggagt ggacccgcca
     1321 cgggagcttt gtcaataagc ccacgcgggg ctggctgcat cccaacgaca aagtcatggg
     1381 acccggggtt tcctacttgg ttcggtacat gggttgtgtg gaggtcctcc agtcaatgcg
     1441 tgccctggac ttcaacaccc ggactcaggt caccagggag gccatcagtc tggtgtgtga
     1501 ggctgtgccg ggtgctaagg gggcgacaag gaggagaaag ccctgtagcc gcccgctcag
     1561 ctctatcctg gggaggagta acctgaaatt tgctggaatg ccaatcactc tcaccgtctc
     1621 caccagcagc ctcaacctca tggccgcaga ctgcaaacag atcatcgcca accaccacat
     1681 gcaatctatc tcatttgcat ccggcgggga tccggacaca gccgagtatg tcgcctatgt
     1741 tgccaaagac cctgtgaatc agagagcctg ccacattctg gagtgtcccg aagggcttgc
     1801 ccaggatgtc atcagcacca ttggccaggc cttcgagttg cgcttcaaac aatacctcag
     1861 gaacccaccc aaactggtca cccctcatga caggatggct ggctttgatg gctcagcatg
     1921 ggatgaggag gaggaagagc cacctgacca tcagtactat aatgacttcc cggggaagga
     1981 accccccttg gggggggtgg tagacatgag gcttcgggaa ggagccgctc caggggctgc
     2041 tcgacccact gcacccaatg cccagacccc cagccacttg ggagctacat tgcctgtagg
     2101 acagcctgtt gggggagatc cagaagtccg caaacagatg ccacctccac caccctgtcc
     2161 aggcagagag ctttttgatg atccctccta tgtcaacgtc cagaacctag acaaggcccg
     2221 gcaagcagtg ggtggtgctg ggccccccaa tcctgctatc aatggcagtg caccccggga
     2281 cctgtttgac atgaagccct tcgaagatgc tcttcgcgtg cctccacctc cccagtcggt
     2341 gtccatggct gagcagctcc gaggggagcc ctggttccat gggaagctga gccggcggga
     2401 ggctgaggca ctgctgcagc tcaatgggga cttcctggta cgagagagca cgaccacacc
     2461 tggccagtat gtgctcactg gcttgcagag tgggcagcct aagcatttgc tactggtgga
     2521 ccctgagggt gtggttcgga ctaaggatca ccgctttgaa agtgtcagtc accttatcag
     2581 ctaccacatg gacaatcact tgcccatcat ctctgcgggc agcgaactgt gtctacagca
     2641 acctgtggag cggaaactgt gaaagccgaa ttctgcagat atccagcaca gtggcggccg
     2701 ctcgagtcta gagggcccgt ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     2761 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     2821 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     2881 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     2941 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3001 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3061 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3121 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3181 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3241 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3301 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3361 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3421 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3481 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3541 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3601 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3661 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3721 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     3781 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     3841 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     3901 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     3961 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4021 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4081 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4141 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4201 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4261 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4321 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4381 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4441 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4501 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4561 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4621 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc
     4681 gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     4741 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     4801 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     4861 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4921 caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc
     4981 gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa
     5041 catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac
     5101 attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca
     5161 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     5221 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     5281 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     5341 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     5401 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     5461 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     5521 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     5581 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     5641 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     5701 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     5761 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     5821 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     5881 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     5941 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     6001 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     6061 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     6121 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     6181 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     6241 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     6301 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     6361 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     6421 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     6481 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     6541 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     6601 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     6661 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     6721 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     6781 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     6841 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     6901 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6961 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     7021 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     7081 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     7141 tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc
     7201 tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag
     7261 tagtgcgcga gcaaaattta agctacaaca aggc
//
