LOCUS       sequence-12154-e        7550 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 7550)
  AUTHORS   Karginov AV, Ding F, Kota P, Dokholyan NV, Hahn KM
  TITLE     Engineered allosteric activation of kinases in living cells.
  JOURNAL   Nat Biotechnol. 2010 Jul;28(7):743-7. doi: 10.1038/nbt.1639. Epub 
            2010 Jun 27.
  PUBMED    20581846
REFERENCE   2  (bases 1 to 7550)
  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: "10.1038/nbt.1639"; 
            journalName: "Nat Biotechnol"; date: "2010-07"; volume: "28"; issue:
            "7"; pages: "743-7"
COMMENT     Sequence Label: pUSE-RapR-Src-myc
FEATURES             Location/Qualifiers
     source          1..7550
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2841..2870
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     complement(3140..3157)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3146..3370
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3416..3844
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3503..3522)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3713..3734
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3853..3873)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3858..4188
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4039..4174
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4101..4120
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4255..5049
                     /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(4309..4328)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4919..4938
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5223..5344
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5260..5279)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5314..5333
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5393..5409)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5393..5409)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5406..5428)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5417..5433
                     /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(5441..5447,5448..5465,5466..5471))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5486..5507
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5624..5641)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5795..6383)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5875..5894)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6554..7345,7346..7414))
                     /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     7177..7196
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7415..7519)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaacgg gccctctaga ctcgagcggc cgccactgtg ctggatcatg ggcagcaaca
      961 agagcaagcc caaggacgcc agccagcggc gccgcagcct ggagccctcg gaaaacgtgc
     1021 acggggcagg gggcgccttc ccggcctcac agacaccgag caagcccgcc tccgccgacg
     1081 gccaccgcgg gcccagcgcc gccttcgtgc cgcccgcggc cgagcccaag ctcttcggag
     1141 gcttcaactc ctcggacacc gtcacctccc cgcagagggc ggggcctctg gcaggtgggg
     1201 tgaccacctt tgtggccctc tatgactatg agtcacggac agagactgac ctgtccttca
     1261 agaaagggga gcggctgcag attgttaaca acacggaggg agactggtgg ctggcacact
     1321 cgctgagcac gggacagacc ggttacatcc ccagcaacta tgtggcgccc tccgactcca
     1381 tccaggctga ggagtggtac tttggcaaga tcactagacg ggaatcagag cggctgctgc
     1441 tcaacgccga gaacccgaga gggaccttcc tcgtgaggga gagtgagacc acaaaaggtg
     1501 cctactgcct ctctgtatcc gacttcgaca atgccaaggg cctaaatgtg aaacactaca
     1561 agatccgcaa gctggacagc ggcggtttct acatcacctc ccgcacccag ttcaacagcc
     1621 tgcagcagct cgtggcttac tactccaaac atgctgatgg cctgtgtcac cgcctcacta
     1681 ccgtatgtcc cacatccaag cctcagaccc agggattggc caaggatgcg tgggagatcc
     1741 cccgggagtc cctgcggctg gaggtcaagc tgggccaggg ttgcttcgga gaggtgtgga
     1801 tggggacctg gaacggacca ggtacctgcg tggtgcacta caccgggatg cttgaagatg
     1861 gaaagaaatt tgattcctcc cgggacagaa acaagccctt taagtttatg ctaggcaagc
     1921 aggaggtgat ccgaggctgg gaagaagggg ttgcccagat gagtgtgggt cagagagcca
     1981 aactgactat atctccagat tatgcctatg gtgccactgg gcacccaggc atcatcccac
     2041 cacatgccac tctcgtcttc gatgtggagc ttctaaaact ggaaggtccc ggaaccacga
     2101 gggttgccat caaaactctg aagccaggca ccatgtcccc agaggccttc ctgcaggagg
     2161 cccaagtcat gaagaaactg aggcacgaga aactggtgca gctgtatgct gtggtgtcgg
     2221 aagaacccat ttacattgtg acagagtaca tgaacaaggg gagtctgctg gactttctca
     2281 agggggaaac gggcaaatat ttgcggctac cccagctggt ggacatgtct gctcagatcg
     2341 cttcaggcat ggcctatgtg gagcggatga actatgtgca ccgggacctt cgagccgcca
     2401 atatcctagt aggggagaac ctggtgtgca aagtggccga ctttgggttg gcccggctca
     2461 tagaagacaa cgaatacaca gcccggcaag gtgccaaatt ccccatcaag tggaccgccc
     2521 ctgaagctgc tctgtacggc aggttcacca tcaagtcgga tgtgtggtcc tttgggattc
     2581 tgctgaccga gctcaccact aagggaagag tgccctatcc tgggatggtg aaccgtgagg
     2641 ttctggacca ggtggagcgg ggctaccgga tgccttgtcc ccccgagtgc cccgagtccc
     2701 tgcatgacct tatgtgccag tgctggcgga aggagcccga ggagcggccc accttcgagt
     2761 acctgcaggc cttcctggaa gactacttta cgtccactga gccacagtac cagcccgggg
     2821 agaacctagg cggaccaggg gagcagaagc tgatctccga ggaggacctg tgaggactgt
     2881 gtgcgcagaa ctgtcacctc taggcttccg ggtgttgcct ccgatgcacc tgcctccctt
     2941 catcctctct ctggggctgc gttgcctcgg caatgcccct tttggagcat ggaagggctt
     3001 cagggatggg gcagcctgga aggacaggtg gaggctggta cagtgactgt cccatgcctg
     3061 ccatgatctg cagaattcca ccacactgga ctagtggatc cgagctcggt accaagctta
     3121 agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     3181 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     3241 aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg
     3301 gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg
     3361 ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg
     3421 ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca
     3481 cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc
     3541 gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct
     3601 ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg
     3661 ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc
     3721 ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg
     3781 attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg
     3841 aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca
     3901 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3961 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     4021 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     4081 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     4141 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     4201 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     4261 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     4321 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     4381 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     4441 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     4501 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     4561 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     4621 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     4681 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     4741 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     4801 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4861 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4921 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4981 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     5041 ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat
     5101 cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc
     5161 gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc
     5221 ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     5281 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     5341 cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca tggtcatagc
     5401 tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca
     5461 taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct
     5521 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     5581 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5641 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5701 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5761 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5821 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5881 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5941 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa tgctcacgct
     6001 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     6061 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6121 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6181 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     6241 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6301 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6361 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6421 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6481 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6541 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6601 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6661 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6721 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6781 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6841 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6901 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6961 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     7021 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     7081 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7141 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7201 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7261 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7321 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7381 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7441 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7501 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
