LOCUS       sequence-61242-e        7140 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 7140)
  AUTHORS   Ogura M, Yamaki J, Homma MK, Homma Y
  TITLE     Mitochondrial c-Src regulates cell survival through phosphorylation 
            of respiratory chain components.
  JOURNAL   Biochem J. 2012 Oct 15;447(2):281-9.
  PUBMED    22823520
REFERENCE   2  (bases 1 to 7140)
  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: "Biochem 
            J."; date: "2012-10-15"; volume: "447(2)"; pages: "281-9"
COMMENT     Sequence Label: pcDNA3-MTS-WT-c-Src-FLAG
FEATURES             Location/Qualifiers
     source          1..7140
                     /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             2659..2682
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     promoter        complement(2692..2710)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2693..2710)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2730..2747)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2736..2960
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3006..3434
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3093..3112)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3303..3324
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3443..3463)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3448..3778
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3629..3764
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3691..3710
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3845..4639
                     /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(3899..3918)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4509..4528
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4813..4934
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4850..4869)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4904..4923
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4983..4999)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4983..4999)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4996..5018)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5007..5023
                     /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(5031..5037,5038..5055,5056..5061))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5076..5097
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5214..5231)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5385..5973)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5465..5484)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6144..6935,6936..7004))
                     /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     6767..6786
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7005..7109)
                     /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 gcttgccacc
      901 atgcaggcgg ctcggatggc cgcgagcttg gggcggcagc tgctgaggct cgggggcgga
      961 agctcgcggc tcacggcgct cctggggcag ccccggcccg gccctgcccg gcggccctat
     1021 gccgggggta ccgagctcgg atctatgggt agcaacaaga gcaagcccaa ggatgccagc
     1081 cagcggcgcc gcagcctgga gcccgccgag aacgtgcacg gcgctggcgg gggcgctttc
     1141 cccgcctcgc agacccccag caagccagcc tcggccgacg gccaccgcgg ccccagcgcg
     1201 gccttcgccc ccgcggccgc cgagcccaag ctgttcggag gcttcaactc ctcggacacc
     1261 gtcacctccc cgcagagggc gggcccgctg gccggtggag tgaccacctt tgtggccctc
     1321 tatgactatg agtctaggac ggagacagac ctgtccttca agaaaggcga gcggctccag
     1381 attgtcaaca acacagaggg agactggtgg ctggcccact cgctcagcac aggacagaca
     1441 ggctacatcc ccagcaacta cgtggcgccc tccgactcca tccaggctga ggagtggtat
     1501 tttggcaaga tcaccagacg ggagtcagag cggttactgc tcaatgcaga gaacccgaga
     1561 gggaccttcc tcgtgcgaga aagtgagacc acgaaaggtg cctactgcct ctcagtgtct
     1621 gacttcgaca acgccaaggg cctcaacgtg aagcactaca agatccgcaa gctggacagc
     1681 ggcggcttct acatcacctc ccgcacccag ttcaacagcc tgcagcagct ggtggcctac
     1741 tactccaaac acgccgatgg cctgtgccac cgcctcacca ccgtgtgccc cacgtccaag
     1801 ccgcagactc agggcctggc caaggatgcc tgggagatcc ctcgggagtc gctgcggctg
     1861 gaggtcaagc tgggccaggg ctgctttggc gaggtgtgga tggggacctg gaacggtacc
     1921 accagggtgg ccatcaaaac cctgaagcct ggcacgatgt ctccagaggc cttcctgcag
     1981 gaggcccagg tcatgaagaa gctgaggcat gagaagctgg tgcagttgta tgctgtggtt
     2041 tcagaggagc ccatttacat cgtcacggag tacatgagca aggggagttt gctggacttt
     2101 ctcaaggggg agacaggcaa gtacctgcgg ctgcctcagc tggtggacat ggctgctcag
     2161 atcgcctcag gcatggcgta cgtggagcgg atgaactacg tccaccggga ccttcgtgca
     2221 gccaacatcc tggtgggaga gaacctggtg tgcaaagtgg ccgactttgg gctggctcgg
     2281 ctcattgaag acaatgagta cacggcgcgg caaggtgcca aattccccat caagtggacg
     2341 gctccagaag ctgccctcta tggccgcttc accatcaagt cggacgtgtg gtccttcggg
     2401 atcctgctga ctgagctcac cacaaaggga cgggtgccct accctgggat ggtgaaccgc
     2461 gaggtgctgg accaggtgga gcggggctac cggatgccct gcccgccgga gtgtcccgag
     2521 tccctgcacg acctcatgtg ccagtgctgg cggaaggagc ctgaggagcg gcccaccttc
     2581 gagtacctgc aggccttcct ggaggactac ttcacgtcca ccgagcccca gtaccagccc
     2641 ggggagaacc tcctcgagga ctacaaggat gacgatgaca aggggcccta ttctatagtg
     2701 tcacctaaat gctagagctc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     2761 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     2821 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     2881 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     2941 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     3001 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     3061 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     3121 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg ggcatccctt tagggttccg
     3181 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     3241 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     3301 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     3361 tttataaggg attttgggga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     3421 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3481 tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     3541 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3601 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     3661 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     3721 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     3781 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     3841 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     3901 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     3961 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     4021 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     4081 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     4141 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     4201 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     4261 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     4321 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     4381 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     4441 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     4501 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     4561 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     4621 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     4681 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     4741 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     4801 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4861 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4921 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     4981 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     5041 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     5101 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     5161 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     5221 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5281 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5341 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5401 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5461 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5521 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     5581 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5641 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     5701 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     5761 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     5821 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     5881 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     5941 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     6001 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     6061 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     6121 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     6181 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     6241 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     6301 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     6361 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     6421 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     6481 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     6541 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     6601 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     6661 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     6721 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     6781 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     6841 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     6901 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     6961 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     7021 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     7081 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
