LOCUS       sequence-99586-e        8091 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP-tagged ORF clone of Homo sapiens neuro-oncological ventral 
            antigen 1 (NOVA1), transcript variant 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8091)
  TITLE     Charlet-Berguerand Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 8091)
  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            "Unpublished"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: NOVA1 eGFP
FEATURES             Location/Qualifiers
     source          1..8091
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     151..173
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     166..182
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     enhancer        343..722
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        723..926
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     877..897
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     952..971
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        952..970
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(2571..2573,2574..3287)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     CDS             join(2571..2573,2574..3284)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     primer_bind     complement(2613..2634)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2874..2893)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3221..3242
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3299..3315)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    3326..3948
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3525..3544)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3972..3992)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3977..4306
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4157..4292
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4373..5167
                     /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(4427..4446)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5037..5056
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5341..5462
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5378..5397)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5432..5451
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5511..5527)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5524..5546)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5535..5551
                     /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(5559..5565,5566..5583,5584..5589))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5604..5625
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5742..5759)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5913..6498)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5993..6012)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6669..7460,7461..7529))
                     /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     7292..7311
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7530..7634)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      7661..8089
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7748..7767)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7958..7979
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 aacaaaatat taacgcttac aatttccatt cgccattcag gctgcgcaac tgttgggaag
       61 ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa
      121 ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca
      181 gtgccaagct gatctataca ttgaatcaat attggcaatt agccatatta gtcattggtt
      241 atatagcata aatcaatatt ggctattggc cattgcatac gttgtatcta tatcataata
      301 tgtacattta tattggctca tgtccaatat gaccgccatg ttgacattga ttattgacta
      361 gttattaata gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg
      421 ttacataact tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga
      481 cgtcaataat gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat
      541 gggtggagta tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa
      601 gtccgccccc tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca
      661 tgaccttacg ggactttcct acttggcagt acatctacgt attagtcatc gctattacca
      721 tggtgatgcg gttttggcag tacaccaatg ggcgtggata gcggtttgac tcacggggat
      781 ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg
      841 actttccaaa atgtcgtaat aaccccgccc cgttgacgca aatgggcggt aggcgtgtac
      901 ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagaatttt gtaatacgac
      961 tcactatagg gcggccggga attcgtcgac tggatccggt accgaggaga tctgccgccg
     1021 cgatcgccat gatggcggca gctcccatcc agcagaacgg gacccacact ggggttccca
     1081 tagacctgga cccgccggac tcgcggaaaa ggccgctgga agccccccct gaagccggca
     1141 gcaccaagag gaccaatacg ggcgaagacg gccagtattt tctaaaggtt ctcataccta
     1201 gttatgctgc tggatctata attgggaagg gaggacagac aattgttcag ttgcaaaaag
     1261 aaactggagc caccatcaag ctgtctaagt ccaaagattt ttacccaggt actactgagc
     1321 gagtgtgctt gatccaggga acggttgaag cactgaatgc agttcatgga ttcattgcag
     1381 aaaaaattcg agaaatgccc caaaatgtgg ccaagacaga accagtcagc attctacaac
     1441 cccagaccac cgttaatcca gatcgcatca aacaaacatt gccatcttcc ccaactacca
     1501 ccaagtcctc tccatctgat cccatgacca cctccagagc taatcaggta aagattatag
     1561 ttcccaacag cacagcaggt ctgataatag ggaagggagg tgctactgtg aaggctgtaa
     1621 tggagcagtc aggggcttgg gtgcagcttt cccagaaacc tgatgggatc aacttgcaag
     1681 agagggttgt cactgtgagt ggagaacctg aacaaaaccg aaaagctgtt gaacttatca
     1741 tccagaagat acaagaggat ccacaaagtg gcagctgtct caatatcagt tatgccaatg
     1801 tgacaggtcc agtggcaaat tccaatccaa ccggatctcc ttatgcaaac actgctgaag
     1861 tgttaccaac tgctgcagca gctgcagggc tattaggaca tgctaacctt gctggcgttg
     1921 cagcctttcc agcagtttta tctggcttca caggcaatga cctggtggcc atcacctctg
     1981 cacttaatac attagccagc tatggatata atctcaacac tttaggttta ggtctcagtc
     2041 aagcagcagc aacaggggct ttggctgcag cagctgccag tgccaaccca gcagcagcag
     2101 cagccaattt attggccacc tatgccagtg aagcctcagc cagtggcagc acagctggtg
     2161 gtacggcggg gacatttgca ttaggtagcc tggctgctgc tactgctgca accaatggat
     2221 attttggagc tgcttctccc ctagctgcca gtgccattct aggaacagaa aagtccacag
     2281 atggatccaa ggatgtagtt gaaatagcag tgccagaaaa cttagttggt gcaatacttg
     2341 gcaaaggagg gaaaacatta gtggaatacc aggagttgac tggtgcaagg atacagatct
     2401 ccaaaaaagg agaattcgta cctggcacaa ggaatcggaa ggtaaccatt actggaacac
     2461 cagctgcaac acaggctgct caatatttaa ttacacaaag gatcacatat gagcaaggag
     2521 ttcgggctgc caatcctcag aaagtgggta cgcgtacgcg gccgctcgag atgagcaagg
     2581 gcgaggagct gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg
     2641 gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc
     2701 tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc
     2761 tgacctacgg cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct
     2821 tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg
     2881 gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg
     2941 agctgaaggg catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca
     3001 actacaacag ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga
     3061 acttcaagat ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc
     3121 agaacacccc catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc
     3181 agtccgccct gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg
     3241 tgaccgccgc cgggatcact ctcggcatgg acgagctgta caagtaaggc cggccgcggt
     3301 catagctgtt tcctgaacag atcccgggtg gcatccctgt gacccctccc cagtgcctct
     3361 cctggccctg gaagttgcca ctccagtgcc caccagcctt gtcctaataa aattaagttg
     3421 catcattttg tctgactagg tgtccttcta taatattatg gggtggaggg gggtggtatg
     3481 gagcaagggg caagttggga agacaacctg tagggcctgc ggggtctatt gggaaccaag
     3541 ctggagtgca gtggcacaat cttggctcac tgcaatctcc gcctcctggg ttcaagcgat
     3601 tctcctgcct cagcctcccg agttgttggg attccaggca tgcatgacca ggctcagcta
     3661 atttttgttt ttttggtaga gacggggttt caccatattg gccaggctgg tctccaactc
     3721 ctaatctcag gtgatctacc caccttggcc tcccaaattg ctgggattac aggcgtgaac
     3781 cactgctccc ttccctgtcc ttctgatttt aaaataacta taccagcagg aggacgtcca
     3841 gacacagcat aggctacctg gccatgccca accggtggga catttgagtt gcttgcttgg
     3901 cactgtcctc tcatgcgttg ggtccactca gtagatgcct gttgaattgg gtacgcggcc
     3961 agcttggctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4021 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4081 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4141 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4201 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4261 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc
     4321 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga
     4381 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga
     4441 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg
     4501 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga
     4561 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt
     4621 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct
     4681 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct
     4741 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg
     4801 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca
     4861 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga
     4921 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt
     4981 ttctggattc atcgactgtg gccggctggg tgtggccgac cgctatcagg acatagcgtt
     5041 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct
     5101 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt
     5161 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca
     5221 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg
     5281 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc
     5341 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     5401 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     5461 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg
     5521 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     5581 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     5641 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     5701 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg
     5761 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     5821 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     5881 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5941 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     6001 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     6061 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     6121 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     6181 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     6241 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     6301 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     6361 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     6421 tccggcaaac aaaccaccgc tggtagcggt ttttttgttt gcaagcagca gattacgcgc
     6481 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     6541 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     6601 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     6661 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     6721 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     6781 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     6841 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     6901 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     6961 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     7021 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     7081 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     7141 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     7201 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     7261 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     7321 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     7381 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     7441 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     7501 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     7561 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     7621 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     7681 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     7741 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     7801 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     7861 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     7921 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     7981 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     8041 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt t
//
