LOCUS       sequence-60974-e        6975 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6975)
  AUTHORS   Loson OC, Song Z, Chen H, Chan DC
  TITLE     Fis1, Mff, MiD49 and MiD51 mediate Drp1 recruitment in mitochondrial
            fission.
  JOURNAL   Mol Biol Cell. 2013 Jan 2.
  PUBMED    23283981
REFERENCE   2  (bases 1 to 6975)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Cell. 2013 Jan 2."
COMMENT     Sequence Label: pcDNA3.1(-) MID51 4xMyc Hisx6
FEATURES             Location/Qualifiers
     source          1..6975
                     /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"
     regulatory      927..936
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2349..2378
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2385..2414
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2421..2450
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2463..2492
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2508..2525
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2569..2586)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2575..2799
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2845..3273
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2932..2951)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3142..3163
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3282..3302)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3287..3616
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3467..3602
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3529..3548
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3683..4477
                     /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(3737..3756)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4347..4366
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4651..4772
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4688..4707)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4742..4761
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4821..4837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4821..4837)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4834..4856)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4845..4861
                     /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(4869..4875,4876..4893,4894..4899))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4914..4935
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5052..5069)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5223..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5303..5322)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5979..6770,6771..6839))
                     /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     6602..6621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6840..6944)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct 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 atgggtggag 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 ctcgaggcca ccatggcagg cgctggtgag cgcaaaggga
      961 agaaggatga caatggcatt ggtacggcca tcgattttgt gctctccaat gcccggctgg
     1021 tgctgggtgt gggtggagca gctatgttgg gcattgccac actggcagtt aagcggatgt
     1081 atgaccgggc aatcagtgcc cctaccagcc ctacccgcct aagccattca gggaagagga
     1141 gctgggagga gccaaactgg atgggctctc ctcgcctatt gaacaaggac atgaaggcag
     1201 gcctgagccg gtccctgcag acccttccca cagactcttc tgcctttgac acagatacat
     1261 tctgcccacc ccggcccaaa ccattggcca ggaggggcca ggtagacttg aagaagtcac
     1321 gactccgcat gtccctgcag gagaaacttc tttcttacta ccggaaccgg gcagccatcc
     1381 ctgctggaga gcaggctcgg gccaagcaag ctgcggtgga catatgtgct gagctccgga
     1441 gcttcctgcg ggccaagctg cctgatatgc cacttcggga catgtacttg agtggcagcc
     1501 tctatgatga tctgcaggtg gtgacggctg accatatcca actcattgtc ccccttgtgc
     1561 tggagcagaa cctgtggtca tgcatacctg gggaggacac catcatgaat gtccctggtt
     1621 tcttcctggt tcgtcgtgag aacccagagt actttcctcg tggtagcagt tattgggacc
     1681 gatgtgttgt aggtggctac ctctccccaa agacagtggc agacaccttt gagaaagtgg
     1741 tggcaggctc catcaactgg ccagccatag ggtccctctt agactatgtg atccgaccag
     1801 caccaccccc agaggccctg acactagaag ttcagtatga gaaggacaaa catcttgtca
     1861 ttgacttcct gccatcagta acccttggtg acactgtctt ggtggccaga ccacaccggt
     1921 tagctcagta tgacaacctg tggcggctga gccttcgccc tgctgagaca gcacgcttac
     1981 gggctttgga ccaggcggac tccggctgcc ggtctctttg cctcaagatc ctcaaggcca
     2041 tatgcaagtc cactccagct ctgggccacc tcactgccag ccagctaacc aacgtcatcc
     2101 tccacttggc ccaggaggag gctgactggt ctccagacat gttggctgac cggtttctac
     2161 aggccctgag gggactcatc agctacttgg aggctggggt cttgcccagt gccctgaacc
     2221 ctaaggtgaa cctatttgca gagctcaccc ctcaagaaat agacgagttg ggatataccc
     2281 tctactgctc gctgtctgag ccggaggtgc tgctgcagac gggatccgag ctcggtacca
     2341 agcttggaga acaaaagttg atttctgaag aagatttgaa cggtgaacaa aagctaatct
     2401 ccgaggaaga cttgaacggt gaacaaaaat taatctcaga agaagacttg aagcttgggc
     2461 ccgaacaaaa actcatctca gaagaggatc tgaatagcgc cgtcgaccat catcatcatc
     2521 atcattgagt ttaaacggtc tccagcttaa gtttaaaccg ctgatcagcc tcgactgtgc
     2581 cttctagttg ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag
     2641 gtgccactcc cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta
     2701 ggtgtcattc tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag
     2761 acaatagcag gcatgctggg gatgcggtgg gctctatggc ttctgaggcg gaaagaacca
     2821 gctggggctc tagggggtat ccccacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2881 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2941 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     3001 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     3061 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     3121 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     3181 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     3241 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     3301 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3361 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     3421 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     3481 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     3541 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     3601 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcaaga
     3661 gacaggatga ggatcgtttc gcatgattga acaagatgga ttgcacgcag gttctccggc
     3721 cgcttgggtg gagaggctat tcggctatga ctgggcacaa cagacaatcg gctgctctga
     3781 tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt ctttttgtca agaccgacct
     3841 gtccggtgcc ctgaatgaac tgcaggacga ggcagcgcgg ctatcgtggc tggccacgac
     3901 gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa gcgggaaggg actggctgct
     3961 attgggcgaa gtgccggggc aggatctcct gtcatctcac cttgctcctg ccgagaaagt
     4021 atccatcatg gctgatgcaa tgcggcggct gcatacgctt gatccggcta cctgcccatt
     4081 cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact cggatggaag ccggtcttgt
     4141 cgatcaggat gatctggacg aagagcatca ggggctcgcg ccagccgaac tgttcgccag
     4201 gctcaaggcg cgcatgcccg acggcgagga tctcgtcgtg acccatggcg atgcctgctt
     4261 gccgaatatc atggtggaaa atggccgctt ttctggattc atcgactgtg gccggctggg
     4321 tgtggcggac cgctatcagg acatagcgtt ggctacccgt gatattgctg aagagcttgg
     4381 cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc gccgctcccg attcgcagcg
     4441 catcgccttc tatcgccttc ttgacgagtt cttctgagcg ggactctggg gttcgaaatg
     4501 accgaccaag cgacgcccaa cctgccatca cgagatttcg attccaccgc cgccttctat
     4561 gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct ccagcgcggg
     4621 gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta taatggttac
     4681 aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt
     4741 tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc gacctctagc
     4801 tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     4861 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     4921 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     4981 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     5041 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     5101 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     5161 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     5221 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     5281 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     5341 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     5401 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     5461 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     5521 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     5581 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     5641 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5701 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6961 gtgccacctg acgtc
//
