LOCUS       sequence-379647-        7599 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Overexpression of Nitric Oxide Associated 1 protein (NOA1), tagged 
            with V5..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7599)
  AUTHORS   Scorrano L
  TITLE     Luca Scorrano plasmids
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 7599)
  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: 
            "unpublished"
COMMENT     Sequence Label: pCDNA3.2/V5-DEST-Noa1
FEATURES             Location/Qualifiers
     source          1..7599
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(150..738)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(230..249)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(909..1700,1701..1769))
                     /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     1532..1551
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1770..1874)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(1949..1968)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        2140..2519
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2520..2723
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2674..2694
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2768..2787
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2768..2786
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2816..2840
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(4944..4968)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             4994..5035
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(5075..5094)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5119..5167
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5369..5797
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5456..5475)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5666..5687
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5806..5826)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5811..6140
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5991..6126
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6053..6072
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             6207..7001
                     /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(6261..6280)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6871..6890
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    7177..7298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7214..7233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7268..7287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(7347..7363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7347..7363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7360..7382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7371..7387
                     /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(7395..7401,7402..7419,7420..7425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7440..7461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(7578..7595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
       61 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
      121 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
      181 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
      241 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
      301 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
      361 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
      421 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
      481 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
      541 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
      601 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
      661 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
      721 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
      781 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
      841 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
      901 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
      961 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     1021 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     1081 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     1141 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     1201 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     1261 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     1321 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     1381 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     1441 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     1501 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     1561 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     1621 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     1681 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     1741 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     1801 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     1861 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg atcgggagat
     1921 ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc
     1981 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
     2041 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
     2101 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
     2161 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
     2221 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
     2281 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
     2341 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
     2401 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
     2461 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
     2521 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
     2581 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
     2641 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
     2701 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
     2761 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagttaagc tatcaacaag
     2821 tttgtacaaa aaagcaggct ccaccatgct gcccgctcgc ctaccgttca ggctgctgag
     2881 ccttttcctt cgtggatccg ctcccacggc agcgcgccat ggcctccggg agccgctcct
     2941 ggagaggagg tgcgctgccg cctcctcctt ccagcactca tcgagtctgg gacgcgagct
     3001 tccttatgac cccgtggaca cggagggctt tggagaaggt ggtgacatgc aggagcgttt
     3061 tctgttcccg gagtacatcc tggatccgga gccgcaaccc acccgcgaaa agcagctgca
     3121 ggagctccag caacagcagg aggaggagga gcgacagagg cagcagcggc gggaggagcg
     3181 gcgacagcaa aacctacggg ccaggtcccg ggagcacccg gtcgtggggc acccggaccc
     3241 ggcattgccg cccagcggcg tgaactgctc gggctgcggg gcagagctgc actgccagga
     3301 cgccggagtg cccggctacc tgccccgaga gaagttcctc cgcacggcgg aggcagacgg
     3361 cgggctggca cggaccgtgt gccagcgctg ctggctgctg tcgcaccacc ggcgcgctct
     3421 acgcctgcag gtgagccgcg agcagtacct ggagctggtg agcgccgcgt tgcggcggcc
     3481 cggcccctcc ctggtgctct acatggtgga cctgctggac ctgcccgacg ccctgctgcc
     3541 cgacttgccc gcgctggtgg gccccaagca gctgatcgtg ctgggaaaca aagtggacct
     3601 cctgccccag gatgctcctg gctaccggca gaggctgcgg gagcgactgt gggaggactg
     3661 tgcccgcgcc gggctcctgc tggcccctgg ccaccaaggg ccacagcgcc ccgtcaagga
     3721 cgagccacag gacggggaga atccgaatcc gccgaactgg tcccgcacag tggtcaggga
     3781 cgtgcggctg atcagcgcca agaccggcta tggagtggaa gagttgatct ctgcccttca
     3841 gcgctcctgg cgctaccgtg gggacgtcta cttagtgggc gccaccaacg ccggcaaatc
     3901 cactctcttt aacacgctcc tggagtccga ttactgcact gccaagggct ccgaggccat
     3961 cgacagagcc accatctccc cttggccagg tactacatta aaccttctga agtttcctat
     4021 ttgcaaccca actccttaca gaatgtttaa aaggcatcaa agacttaaaa aagattcaac
     4081 tcaagctgaa gaagatctta gtgagcaaga acaaaatcag cttaatgtcc tcaaaaagca
     4141 tggttatgtc gtaggaagag ttggaaggac attcttgtat tcagaagaac agaaggataa
     4201 cattcccttt gagtttgatg ctgattcact tgcctttgac atggaaaatg accctgttat
     4261 gggtacacac aaatccacca aacaagtaga attgactgca caagatgtga aagatgccca
     4321 ctggttttat gacacccctg gaattacaaa agaaaattgt attttaaatc ttctaacaga
     4381 aaaagaagta aatattgttt tgccaacaca gtccattgtt ccaagaactt ttgtgcttaa
     4441 accaggaatg gttctgtttt tgggtgctat aggccgcata gatttcctgc agggaaatca
     4501 gtcagcttgg tttacagtcg tggcttccaa catcctccct gtgcatatca cctccttgga
     4561 cagggcagac gctctgtatc agaagcatgc aggtcatacg ttactccaga ttccaatggg
     4621 tggaaaagaa cgaatggcag gatttcctcc tcttgttgct gaagacatta tgttaaaaga
     4681 aggactgggg gcatctgaag cagtggccga catcaagttt tcctctgcag gttgggtttc
     4741 agtaacacct aattttaagg acagactgca tctccgaggc tatacacctg aaggaacagt
     4801 tttgaccgtc cggccccctc tcttgccata tattgttaac atcaaaggac agcgcatcaa
     4861 gaaaagtgtg gcctataaaa ccaagaagcc tccttccctt atgtacaacg tgaggaagaa
     4921 gaaaggaaag ataaatgtat tggacccagc tttcttgtac aaagtggttg atctagaggg
     4981 cccgcggttc gaaggtaagc ctatccctaa ccctctcctc ggtctcgatt ctacgcgtac
     5041 cggttagtaa tgagtttaaa cgggggaggc taactgaaac acggaaggag acaataccgg
     5101 aaggaacccg cgctatgacg gcaataaaaa gacagaataa aacgcacggg tgttgggtcg
     5161 tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga
     5221 ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg
     5281 ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat agcagatctg
     5341 cgcagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     5401 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     5461 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     5521 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     5581 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     5641 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     5701 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     5761 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     5821 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     5881 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5941 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     6001 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     6061 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     6121 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     6181 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     6241 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     6301 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     6361 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     6421 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     6481 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     6541 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     6601 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     6661 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     6721 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     6781 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     6841 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     6901 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     6961 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcgc
     7021 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     7081 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     7141 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     7201 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     7261 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     7321 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     7381 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     7441 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     7501 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     7561 gggcgctctt ccgcttcctc gctcactgac tcgctgcgc
//
