LOCUS       sequence-10629-e        8056 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 8056)
  AUTHORS   Chen H, Detmer SA, Ewald AJ, Griffin EE, Fraser SE, Chan DC
  TITLE     Mitofusins Mfn1 and Mfn2 coordinately regulate mitochondrial fusion 
            and are essential for embryonic development.
  JOURNAL   J Cell Biol. 2003 Jan 20. 160(2):189-200.
  PUBMED    12527753
REFERENCE   2  (bases 1 to 8056)
  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: "J Cell 
            Biol. 2003 Jan 20. 160(2):189-200."
COMMENT     Sequence Label: Mfn1-Myc
FEATURES             Location/Qualifiers
     source          1..8056
                     /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             3165..3194
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3201..3230
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3237..3266
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3285..3314
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3321..3350
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3357..3386
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3405..3434
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3441..3470
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3477..3506
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3540..3569
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3585..3602
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3646..3663)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3652..3876
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3922..4350
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4009..4028)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4219..4240
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4359..4379)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4364..4694
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4545..4680
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4607..4626
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4761..5555
                     /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(4815..4834)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5425..5444
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5729..5850
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5766..5785)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5820..5839
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5899..5915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5899..5915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5912..5934)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5923..5939
                     /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(5947..5953,5954..5971,5972..5977))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5992..6013
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6130..6147)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6301..6889)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6381..6400)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7060..7851,7852..7920))
                     /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     7683..7702
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7921..8025)
                     /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 gccctctagg tgcataatgg cagaaacggt atctccactg aagcacttcg
      961 tgctggcaaa gaaagccatc actgcaatct tcggccagtt actggagttt gttactgagg
     1021 gctcacattt tgttgaagca acatacagaa atccagaact tgatcgaata gcatccgagg
     1081 atgatctggt ggaaatacag ggctacagaa acaagcttgc tgtcattggg gaggtgctgt
     1141 ctcggagaca tatgaaggtg gcattttttg gcaggacaag tagtggcaag agctctgtca
     1201 tcaatgcaat gctgtgggat aaagtcctcc ccagcgggat tggtcacaca accaactgct
     1261 tcctgagtgt cgaggggacc gatggagata aagcctacct tatgaccgaa gggtcagatg
     1321 aaaagaaaag tgtgaagact gttaatcagc tggcccatgc cctccatatg gataaagact
     1381 tgaaagctgg ctgtcttgtg catgtatttt ggcccaaagc aaaatgtgcc ctcttgagag
     1441 atgacctggt tttagtagac agcccaggta cagatgtcac cacagagctg gatatctgga
     1501 ttgataagtt ttgccttgat gctgatgtct ttgttttggt tgcaaactcg gaatcaacac
     1561 tgatgaacac ggagaaacat tttttccata aggtgaatga gcggctctcc aagcccaaca
     1621 tcttcattct gaataaccgt tgggatgctt ctgcttcgga gccggagtac atggaggatg
     1681 tgcgcagaca gcacatggag agatgtcttc acttcttggt agaagagctc aaggttgtaa
     1741 gtccgtcgga agctcggaat cggatctttt ttgtttcagc caaggaagtt ctcaactcca
     1801 gaaagcataa agctcagggg atgccagaag gtggtggggc acttgcagaa ggatttcaag
     1861 caagattaca ggagtttcaa aattttgaac aaacttttga ggagtgtatc tcgcagtcag
     1921 cagtgaaaac aaagtttgaa cagcacacta tcagagctaa acagatacta gacactgtga
     1981 aaaacatact ggactcagta aacgtggcag cagcagagaa gagggtttat tcaatggaag
     2041 agagggaaga ccaaatcgat agactggact ttatccgaaa ccagatgaac cttttaacac
     2101 tggatgttaa gaagaagatc aaggaggtca cggaggaggt ggcaaacaag gtttcttgtg
     2161 caatgacaga tgaaatttgt cgactatctg ttttggttga tgagttttgt tctgagtttc
     2221 atcctacccc cagtgtactg aaagtgtata agagtgagtt aaataagcac atagaagatg
     2281 gcatgggaag aaatttggct gatcggtgta ccaatgaagt caatgcctcc attcttcaat
     2341 ctcagcaaga aatcatcgaa aacttgaagc cactacttcc agctggtata cagaataaac
     2401 ttcatacatt aatcccttgc aaaaagtttg acctcagcta tgatctcaat tgccacaagc
     2461 tgtgttcgga ttttcaagag gacattgtgt ttcggttttc cctgggctgg tcttcccttg
     2521 tacatcgatt cctgggttcc acaaatgcac agagggtgct gctcgggctg tcagagccca
     2581 tctttcaggt ccctagatct ttagcttcaa ctcctactgc tccttctaac ccagcagccc
     2641 cggataatgc agcccaggag gagctcatga tcaccctgat cacaggattg gcgtccctca
     2701 cgtcgagaac ctccatgggc atcatcgttg ttgggggcgt gatttggaaa acagtgggct
     2761 ggaaactaat ctctgtcacc ttaagtatgt acggagctct gtacctttat gagaggctga
     2821 cgtggacgac ccgtgcgaaa gagagagcgt ttaagcagca gtttgtaaac tatgcaaccg
     2881 agaagctgca gatgattgtg agcttcacca gtgcaaactg cagccaccaa gtacagcaag
     2941 aaatggccac tacttttgct cgactgtgcc aacaagttga tgttactcag aaacatctgg
     3001 aagaggaaat tgcaagatta tccaaagaga tagaccaact ggagaaaata cagaacaact
     3061 caaagctctt aagaaataaa gctgttcaac ttgaaagtga gctggagaat ttttcgaagc
     3121 agtttctaca cccgagcagt ggagaatccg gatcctctag aggtgaacaa aagttgattt
     3181 ctgaagaaga tttgaacggt gaacaaaagc taatctccga ggaagacttg aacggtgaac
     3241 aaaaattaat ctcagaagaa gacttgaacg gatcctctag aggtgaacaa aagttgattt
     3301 ctgaagaaga tttgaacggt gaacaaaagc taatctccga ggaagacttg aacggtgaac
     3361 aaaaattaat ctcagaagaa gacttgaacg gatcctctag aggtgaacaa aagttgattt
     3421 ctgaagaaga tttgaacggt gaacaaaagc taatctccga ggaagacttg aacggtgaac
     3481 aaaaattaat ctcagaagaa gacttgaacg gatccgagct cggtaccaag cttgggcccg
     3541 aacaaaaact catctcagaa gaggatctga atagcgccgt cgaccatcat catcatcatc
     3601 attgagttta aacggtctcc agcttaagtt taaaccgctg atcagcctcg actgtgcctt
     3661 ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg
     3721 ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt
     3781 gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca
     3841 atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct
     3901 ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg
     3961 tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt
     4021 tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcggggca
     4081 tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg
     4141 gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg
     4201 agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct
     4261 cggtctattc ttttgattta taagggattt tggggatttc ggcctattgg ttaaaaaatg
     4321 agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg
     4381 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     4441 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4501 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4561 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4621 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4681 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4741 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4801 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4861 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4921 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4981 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     5041 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     5101 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     5161 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     5221 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5281 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5341 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5401 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5461 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5521 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5581 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5641 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5701 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5761 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5821 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5881 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5941 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     6001 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     6061 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     6121 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     6181 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6241 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6301 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6361 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6421 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6481 cgtggcgctt tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6541 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6601 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6661 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6721 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     6781 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     6841 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     6901 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     6961 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     7021 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     7081 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     7141 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     7201 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     7261 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     7321 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     7381 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     7441 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     7501 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     7561 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     7621 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     7681 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     7741 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     7801 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     7861 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     7921 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     7981 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     8041 agtgccacct gacgtc
//
