LOCUS       sequence-379657-        9191 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expression of OPA1..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9191)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 9191)
  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: 
            "unpublished"
COMMENT     Sequence Label: pcDNA3.1-Opa1
FEATURES             Location/Qualifiers
     source          1..9191
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(1..589)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(81..100)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(760..1551,1552..1620))
                     /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     1383..1402
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1621..1725)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1800..1819)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        1991..2370
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2371..2574
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2525..2545
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2619..2638
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2619..2637
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6474..6490)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6474..6490)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     promoter        complement(6500..6518)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(6501..6518)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(6538..6555)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6544..6768
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      6814..7242
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6901..6920)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7111..7132
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(7251..7271)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7256..7585
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7436..7571
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7498..7517
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             7652..8446
                     /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(7706..7725)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8316..8335
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    8620..8741
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8657..8676)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8711..8730
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(8790..8806)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8790..8806)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8803..8825)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8814..8830
                     /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(8838..8844,8845..8862,8863..8868))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8883..8904
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(9021..9038)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
       61 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
      121 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
      181 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
      241 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
      301 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
      361 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
      421 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
      481 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
      541 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
      601 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
      661 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
      721 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
      781 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
      841 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
      901 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
      961 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     1021 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     1081 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     1141 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     1201 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     1261 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     1321 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     1381 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     1441 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     1501 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     1561 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     1621 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     1681 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     1741 agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg cactctcagt
     1801 acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg tgtgttggag
     1861 gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt gaccgacaat
     1921 tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt acgggccaga
     1981 tatacgcgtt gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta
     2041 gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc
     2101 tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg
     2161 ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg
     2221 gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa
     2281 tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac
     2341 atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg
     2401 cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg
     2461 agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca
     2521 ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg
     2581 ctaactagag aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag
     2641 acccaagctt ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattct
     2701 gcagatatcc atcacactgg cggccgctcg acggtatcga taagcttgat gggactcggt
     2761 ggctgctcta tgcccttgtt gctgagggcc tcttcctggg tcattgtcgg agcaggaatc
     2821 ggactggggc cgacacgggg ttccccccgt ggccgtctca gcgcctgtgt gtggtctgct
     2881 ctggcgggat gtggcgagca ggtcgggcgg ccgtggcctg tgaagtctgc caatccttag
     2941 tgaaacacag ctctggaata caaagaaacg taccgctcca aaaactccat ctggtttcac
     3001 gaagtattta tcgttcacat catcctgccc tcaagcttca aagaccccaa ctaaggacac
     3061 catttcagca gttctcttct ctaactcacc tttcattaca taaattgaaa ctttctccaa
     3121 ttaaatatgg ctaccagccc cgcaggaact tttggccagc aaggttagct gcaagactct
     3181 taaaacttcg atatatcata ctgggatctg ctgttggagg tggctataca gccaaaaaaa
     3241 ccttcgatga atggaaagat atgataccag accttagtga ctataagtgg attgtgcctg
     3301 actttatatg ggaaattgat gagtatattg atttggagaa aattagaaaa gcccttccca
     3361 gctcagaaga ccttgccagt ttagctcccg acttggacaa gattactgag agcctcagct
     3421 tgttgaagga cttcttcact gcaggttcac ctggagaaac agcatttcga gcaacagatc
     3481 atggatctga aagtgacaag cattacagga aggtgtcaga caaagaaaag attgaccaac
     3541 ttcaagaaga acttctgcat actcagttaa agtatcagag gatcttggag cgcctggaaa
     3601 aggagaacaa agagctgagg aagctggtgc tgcagaagga cgacaaaggc atccaccaca
     3661 ggaagctcaa gaaatctttg attgatatgt attctgaagt tcttgatgtt ctttctgatt
     3721 atgatgccag ttacaataca caagatcacc taccacgggt tgttgtggtt ggagatcaga
     3781 gtgctgggaa aaccagtgtg ctggaaatga ttgctcaggc ccggatcttc ccgagagggt
     3841 ccggcgagat gatgacacgc tctccagtga aggtgactct cagtgaaggc cctcaccatg
     3901 tggccttgtt taaagatagc tctcgggaat ttgatctcac caaggaggaa gatcttgcag
     3961 cattaagaca tgaaatcgaa ctccgaatga ggaaaaatgt gaaagaaggt tgtactgtta
     4021 gccccgagac catatctcta aatgtcaaag gccctgggct gcagaggatg gtgctcgtgg
     4081 acttgcctgg tgtcatcaac actgtgacat caggcatggc tcccgacaca aaggaaacta
     4141 ttttcagtat cagcaaagct tacatgcaga atcctaacgc catcatcctg tgcatccaag
     4201 acggatccgt agatgctgag cgcagtattg ttacagactt ggtcagtcaa atggatcctc
     4261 atggaagaag aaccatattt gttttgacca aagtagacct ggcagaaaaa aatgtagcca
     4321 gtccaagcag gatacaacag ataattgaag gcaagctctt cccaatgaaa gctctgggtt
     4381 attttgctgt cgtaacagga aaaggaaaca gctctgaaag tattgaagct ataagagagt
     4441 atgaagagga attttttcag aattcaaaac tgctaaagac aagcatgcta aaggcacacc
     4501 aggtcaccac gagaaatctc agccttgctg tgtcagactg cttttggaaa atggttcgag
     4561 agtcagttga acaacaggct gatagtttta aagccacgcg ctttaaccta gagacggaat
     4621 ggaagaataa ctacccgcgc ctgcgagagc tcgacaggaa tgaactcttt gaaaaagcta
     4681 aaaatgagat cctcgatgag gtcatcagtc tgagccaggt cacgccaaag cactgggagg
     4741 aaatcctgca gcaatccctg tgggaacgag tgtcaacaca tgtgattgag aacatctacc
     4801 ttccagctgc ccagaccatg aattcgggaa catttaacac cacagtagac atcaagctta
     4861 aacagtggac tgacaagcag cttcctaata aagcagtcga ggttgcctgg gagactctac
     4921 aagaggaatt ttcccgcttc atgacagaac ccaaaggaaa ggaacacgac gacatatttg
     4981 acaaacttaa ggaggctgtg aaggaggaga gtatcaagcg gcacaagtgg aacgactttg
     5041 ccgaggatag cttgagggtt attcagcaca atgctttgga agaccggtcc atatcagata
     5101 agcaacagtg ggatgcagcc atttacttca tggaagaggc gcttcaaggt cgtctcaagg
     5161 atactgaaaa tgctattgaa aacatgattg ggccagactg gaaaaagagg tggatgtact
     5221 ggaagaatcg gacccaagag cagtgtgttc acaacgaaac caagaacgag ttggagaaga
     5281 tgctgaaggt taatgatgag cacccagctt acctggcaag tgatgagatt accacagtcc
     5341 ggaagaacct ggagtctcga ggagtggaag tcgatccaag cttgattaag gatacttggc
     5401 atcaagttta tagaagacat ttcttaaaaa cagctctaaa tcattgtaac ctttgtcgca
     5461 gaggttttta ttactaccag aggcatttta tagattctga gctggaatgc aatgacgtgg
     5521 tcctgttttg gcgaatacag cgcatgctcg ctatcactgc caatacatta aggcagcagc
     5581 ttacaaacac tgaagttagg cgactagaga aaaacgttaa agaggtatta gaagattttg
     5641 cagaagacgg tgagaagaag gttaaattgc tcactggcaa acgagttcag ctggcagaag
     5701 atctcaagaa agttagagaa attcaagaaa agcttgatgc tttcattgaa gctcttcacc
     5761 aggagaagta gactgtgtca aaatcctgct cataatgtaa tcacctctgc gtttatttga
     5821 agaatgaaaa cttgagtctt ttgtcctgcc tcctttcctc ccgttctacc cttttccgtg
     5881 caaaataaag tcacagggta aaagtcatgt acgtgtgttc caggcactta acctgcagct
     5941 gccgtttgag cagaagagcc ctggacgtgg catccacggt ctgtgagctc gtgctgacgg
     6001 cagacattgg gaaaacatca ctggtgtggc cgtgaggctt agtcctcaga gattaagaag
     6061 ttgctctctg agtcgtgttt gtggtggcac tgatgtgatt cactatggct ggtatcacac
     6121 gtacagtggt agctgctctg gaataaaact tacagtaatt ttttaaaagg catcaattgt
     6181 atatgacaag atgacagtta ctgctaaacc atcatgtcta cagtgttctg tattcaggaa
     6241 cgctttgact gtaaacactc agtttatgtt caccacatga aagaaagctg gtttataaac
     6301 agatgcttca gctgggttaa tttatgttgg gcagttcagt gtatattcag tgaaagaaac
     6361 atccaccctc cattgccagc ttagaaactc gccttggaag ccaaccgcag cagcgtgcag
     6421 tccatcgctg cttttgcgcc ctgtatccgt gatcgaattc ctgcagcccg ggggatccac
     6481 tagttctaga gggccctatt ctatagtgtc acctaaatgc tagagctcgc tgatcagcct
     6541 cgactgtgcc ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga
     6601 ccctggaagg tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcgcatt
     6661 gtctgagtag gtgtcattct attctggggg gtggggtggg gcaggacagc aagggggagg
     6721 attgggaaga caatagcagg catgctgggg atgcggtggg ctctatggct tctgaggcgg
     6781 aaagaaccag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     6841 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     6901 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     6961 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     7021 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     7081 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     7141 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     7201 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     7261 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     7321 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     7381 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     7441 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     7501 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     7561 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     7621 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     7681 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     7741 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     7801 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     7861 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     7921 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     7981 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     8041 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     8101 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     8161 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     8221 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     8281 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     8341 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     8401 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     8461 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     8521 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     8581 cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat
     8641 aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg
     8701 cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg
     8761 acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat
     8821 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     8881 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     8941 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     9001 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     9061 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     9121 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     9181 ttgctggcgt t
//
