LOCUS       sequence-60958-e        6948 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 6948)
  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 6948)
  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: "Mol Biol 
            Cell. 2013 Jan 2."
COMMENT     Sequence Label: pcDNA3.1(-) MID49 4xMyc Hisx6
FEATURES             Location/Qualifiers
     source          1..6948
                     /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"
     protein_bind    1107..1118
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             2322..2351
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2358..2387
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2394..2423
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2436..2465
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2481..2498
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2542..2559)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2548..2772
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2818..3246
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2905..2924)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3115..3136
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3255..3275)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3260..3589
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3440..3575
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3502..3521
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3656..4450
                     /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(3710..3729)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4320..4339
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4624..4745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4661..4680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4715..4734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4794..4810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4794..4810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4807..4829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4818..4834
                     /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(4842..4848,4849..4866,4867..4872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4887..4908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5025..5042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5196..5781)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5276..5295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5952..6743,6744..6812))
                     /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     6575..6594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6813..6917)
                     /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 ccatggcaga gttctcccag aagcagagga
      961 aacagagcgg cagcgagggc ctgggcagtg tggtggactt cctcttggct aatgctcgat
     1021 tggtgctagg tgtgggcggg gctgcagtgc tgggcattgc taccctggct gtaaagcggc
     1081 tcatcgacag ggccaccagc cctccagatg aggatgacac caagggggac agctggaagg
     1141 aactgagcct gctgagagcc acatcgcccc aaaagcccca accccctcct gctgccttca
     1201 gccagcccct agctactggg tccccctccc cctctgtgcc agtggagccc acgcccattc
     1261 attctcccac gacacctaag ttcagcacta tagcacccct gtgcctgacg ttccaggaga
     1321 ggctgctggc cttcgagcgc aagcatgtga ttaccccaga agcccacgtg acgttggcga
     1381 aacagctggc tggagacatt gccttggagc tgcaggccta cttgaggagc aagttcccag
     1441 aactgccctt tggtgcactc gtgccgggtg ggcctctcta cgacgggctc caggcaggga
     1501 ctgctgagca cgtgcgcctg ctggcgcctc tggagttgga gccggggcta tggagcctgg
     1561 tacctggcgt ggacactgtg gccagggagc ctcgatgctg ggctgtacgc aggactcagc
     1621 ttgagtttca tccccgtggg tgcagcccat gggaccgctt cctggtgggg ggctacctct
     1681 cctcccgagt cttgctggag ctgctacgga aggccctgtc agcctctgtc aactggccag
     1741 ccattggcag cctgctcggg tgtctgatct ggcccgatgt ggcttcggag gagctgctgc
     1801 tcaaagtgca acacgagtgc ctggagttca ctttagctgt gctcatggta gtccctgggg
     1861 ccagcactga tgaccgcctg ctgctggctt ggcccctaga ggggctggcc agcaatctct
     1921 ggctgcagga tctatatcca gtagagaccg cccgtttgcg ggccctggat gaccaggatg
     1981 ctggtactcg gcggaggttg ttgctgctgc tgtgtggtat ctgtcgtggc catccggctc
     2041 tagtgcggct gggctggagc cacctgactc aggtggttct gcatctgggt gaagaggaag
     2101 tggcctggac tgaggaggcc ttgggggaac ggtttctgca agccctggag tttctggtgg
     2161 gcagcttgga gcaggctagc ctgccctgtc acttcaaccc cagtgtgaac ctcttgggta
     2221 acttccggga agaggagatt gatgacattg gctacgtgct gtacagcggc ctccaggttc
     2281 ctgaaagcct attcggatcc gagctcggta ccaagcttgg agaacaaaag ttgatttctg
     2341 aagaagattt gaacggtgaa caaaagctaa tctccgagga agacttgaac ggtgaacaaa
     2401 aattaatctc agaagaagac ttgaagcttg ggcccgaaca aaaactcatc tcagaagagg
     2461 atctgaatag cgccgtcgac catcatcatc atcatcattg agtttaaacg gtctccagct
     2521 taagtttaaa ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     2581 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     2641 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     2701 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     2761 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     2821 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     2881 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     2941 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3001 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3061 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3121 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     3181 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     3241 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3301 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     3361 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     3421 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     3481 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     3541 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     3601 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     3661 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     3721 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     3781 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     3841 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     3901 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     3961 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4021 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4081 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4141 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     4201 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     4261 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     4321 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     4381 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     4441 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     4501 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     4561 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     4621 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     4681 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     4741 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     4801 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4861 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4921 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4981 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5041 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5101 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     5221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     5281 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     5341 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     5401 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     5461 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     5521 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     5581 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     5641 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     5701 tgatccggca aacaaaccac cgctggtagc ggtttttttg tttgcaagca gcagattacg
     5761 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5821 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5881 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5941 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     6001 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     6061 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     6121 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6181 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6241 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6301 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6361 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6421 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6481 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6541 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6601 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6661 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6721 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6781 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6841 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6901 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
