LOCUS       578#_pHSV_vamp7        6826 bp DNA     circular SYN 24-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6826)
  AUTHORS   Alonso
  TITLE     Direct Submission
  JOURNAL   Exported Nov 24, 2025 from SnapGene 7.2.1
            https://www.snapgene.com
COMMENT     Sequence Label: pHSV_ELGA1
FEATURES             Location/Qualifiers
     source          1..6826
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    186..326
                     /label=bom
                     /note="basis of mobility region from pBR322"
     regulatory      1499..1508
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1505..3523
                     /codon_start=1
                     /product="GFP-Aequorin fusion is a domain of erGAP1 
                     described in Rodriguez-Garcia, PNAS 2014, renamed here as 
                     ELGA, to make it more comprehensive."
                     /label=ELGA1
                     /translation="MAILFAVVARGTTILAKHAWCGGNFLEVTEQILAKIPSENNKLTY
                     SHGNYLFHYICQDRIVYLCITDDDFERSRAFNFLNEIKKRFQTTYGSRAQTALPYAMNS
                     EFSSVLAAQLKHHSENKGLDKVMETQAQVDELKGIMVRNIDLVAQRGERLELLIDKTEN
                     LVDSSVTFKTTSRNLARAMCMKNLKLTIIIIIVSIVFIYIIVSPLCGGFTWPSCVKKKL
                     YDVPDYASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKL
                     PVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGNYKTRAEV
                     KFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKANFKIRHNI
                     EDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGIT
                     HGMDELYKPGTATPATTPTTAPTAGTGKLTSDFDNPRWIGRHKHMFNFLDVNHNGKISL
                     DEMVYKASDIVINNLGATPEQAKRHKDAVEAFFGGAGMKYGVETDWPAYIEGWKKLATD
                     ELEKYAKNEPTLIRIWGDALFDIVAKAQNGAITLDEWKAYTKAAGIIQSSEDCEETFRV
                     CDIDESGQLDVAEMTRQHLGFWYTMDPACEKLYGGAVP"
     misc_feature    1505..2164
                     /label=VAMP
                     /note="VAMP"
     primer_bind     2189..2212
                     /label=Primer 1
     misc_feature    2190..2911
                     /label=GFP
                     /note="GFP (C3 GFP)"
     misc_feature    2912..2956
                     /label=Linker
                     /note="Linker"
     CDS             2957..3523
                     /codon_start=1
                     /label=mutAequorin
                     /note="mut Aequorin"
                     /translation="GKLTSDFDNPRWIGRHKHMFNFLDVNHNGKISLDEMVYKASDIVI
                     NNLGATPEQAKRHKDAVEAFFGGAGMKYGVETDWPAYIEGWKKLATDELEKYAKNEPTL
                     IRIWGDALFDIVAKAQNGAITLDEWKAYTKAAGIIQSSEDCEETFRVCDIDESGQLDVA
                     EMTRQHLGFWYTMDPACEKLYGGAVP"
     misc_feature    3305..3307
                     /label=D117A
                     /note="D117A mutation in Aequorin gene to reduce Ca2+ 
                     affinity"
     misc_feature    3311..3313
                     /label=D119A
                     /note="D119A mutation in Aequorin gene to reduce Ca2+ 
                     affinity"
     misc_feature    3443..3445
                     /label=D163A
                     /note="D163A mutation in Aequorin gene to reduce Ca2+ 
                     affinity"
     polyA_signal    3847..3980
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        5102..5206
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5207..5275,5276..6067)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6238..6826
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      121 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      181 gagcgcctga tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcatatgg
      241 tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac actccgctat
      301 cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct gacgcgccct
      361 gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc tccgggagct
      421 gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagccg gccattatgc
      481 acgaccccgc cccgacgccg gcacgccggg ggcccgtggc cgcggcccgt tggtcgaacc
      541 cccggccccg cccatccgcg ccatctgcca tgggcggggc gcgagggcgg gtgggtccgc
      601 gccccgcccc gcatggcatc tcattaccgc ccgatccggc ggtttccgct tccgttccgc
      661 atgctaacga ggaacgggca gggggcgggg cccgggcccc gacttcccgg ttcggcggta
      721 atgagatacg agccccgcgc gcccgttggc cgtccccggg ccccccggtc ccgcccgccg
      781 gacgccggga ccaacgggac ggcgggcggc ccaagggccg cccgccttgc cgccccccca
      841 ttggccggcg ggcgggaccg ccccaagggg gcggggccgc cgggtaaaag aagtgagaac
      901 gcgaagcgtt cgcacttcgt cccaatatat atatattatt agggcgaagt gcgagcactg
      961 gcgccgtgcc cgactccgcg ccggccccgg gggcgggccc gggcggcggg gggcgggtct
     1021 ctccggcgca cataaaggcc cggcgcgacc gacgcccgca gacggcgccg gccacgaacg
     1081 acgggagcgg ctgcggagca cgcggaccgg gagcgggagt cgcagagggc cgtcggagcg
     1141 gacggcgtcg gcatcgcgac gccccggctc gggatcggga tcgcatcgga aagggacacg
     1201 cggacgcggg ggggaaagac ccgcccaccc cacccacgaa acacagggga cgcaccccgg
     1261 gggcctccga cgacagaaac ccaccggtcc gccttttttg cacgggtaag caccttgggt
     1321 gggcggagga gggggggacg cgggggtgga ggagggggga cgcgggggcg gaggaggggg
     1381 gacgcggggg cggaggaggg gggacgcggg ggcggaggag gggggacgcg ggggcggagg
     1441 agggggctca cccgcgttcg tgccttcccg caggaggaac gtcctcgtcg ataagcttgc
     1501 caccatggcg attctttttg ctgttgttgc cagggggacc actatccttg ccaaacatgc
     1561 ttggtgtgga ggaaacttcc tggaggtgac agagcagatt ctggctaaga taccttctga
     1621 aaataacaaa ctaacgtact cacatggcaa ttatttgttt cattacatct gccaagacag
     1681 gattgtatat ctttgtatca ctgatgatga ttttgaacgt tcccgagcct ttaattttct
     1741 gaatgagata aagaagaggt tccagactac ttacggttca agagcacaga cagcacttcc
     1801 atatgccatg aatagcgagt tctcaagtgt cttagctgca cagctgaagc atcactctga
     1861 gaataagggc ctagacaaag tgatggagac tcaagcccaa gtggatgaac tgaaaggaat
     1921 catggtcaga aacatagatc tggtagctca gcgaggagaa agattggaat tattgattga
     1981 caaaacagaa aatcttgtgg attcttctgt caccttcaaa actaccagca gaaatcttgc
     2041 tcgagccatg tgtatgaaga acctcaagct cactattatc atcatcatcg tatcaattgt
     2101 gttcatctat atcattgttt cacctctctg tggtggattt acatggccaa gctgtgtgaa
     2161 gaaaaagctt tatgatgttc ctgattatgc tagcaaagga gaagaacttt tcactggagt
     2221 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     2281 agagggtgaa ggtgatgcta catacggaaa acttaccctt aaatttattt gcactactgg
     2341 aaaactacct gttccatggc caacacttgt cactactttc tcttatggtg ttcaatgctt
     2401 ttcccgttat ccggatcata tgaaacggca tgactttttc aagagtgcca tgcccgaagg
     2461 ttatgtacag gaacgcacta tatctttcaa agatgacggg aactacaaga cgcgtgctga
     2521 agtcaagttt gaaggtgata cccttgttaa tcgtatcgag ttaaaaggta ttgattttaa
     2581 agaagatgga aacattctcg gacacaaact cgagtacaac tataactcac acaatgtata
     2641 catcacggca gacaaacaaa agaatggaat caaagctaac ttcaaaattc gccacaacat
     2701 tgaagatgga tccgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg
     2761 ccctgtcctt ttaccagaca accattacct gtcgacacaa tctgcccttt cgaaagatcc
     2821 caacgaaaag cgtgaccaca tggtccttct tgagtttgta actgctgctg ggattacaca
     2881 tggcatggat gagctctaca aaccgggtac agcgacccca gcaaccacgc cgaccactgc
     2941 accgaccgcg ggtaccggta aacttacatc agacttcgac aacccaagat ggattggacg
     3001 acacaagcat atgttcaatt tccttgatgt caaccacaat ggaaaaatct ctcttgacga
     3061 gatggtctac aaggcatctg atattgtcat caataacctt ggagcaacac ctgagcaagc
     3121 caaacgacac aaagatgctg tggaagcctt cttcggagga gctggaatga aatatggtgt
     3181 ggaaactgat tggcctgcat atattgaagg atggaaaaaa ttggctactg atgaattgga
     3241 gaaatacgcc aaaaacgaac caaccctcat ccgcatctgg ggtgatgctt tgtttgacat
     3301 cgttgccaaa gcccaaaatg gagccattac actggatgaa tggaaagcat acaccaaagc
     3361 tgctggtatc atccaatcat cagaagattg cgaggaaaca ttcagagtgt gcgatattga
     3421 tgaaagtgga caactcgatg ttgccgagat gacaagacag catctgggat tttggtacac
     3481 catggatcct gcttgcgaaa agctctacgg tggagctgtc ccctaagaat tccagctgag
     3541 cgccggtcgc taccattacc agttggtctg gtgtcaaaaa taataataac cgggcaggcc
     3601 atgtctgccc gtatttcgcg taaggaaatc cattatgtac tatttaaaaa acacaaactt
     3661 ttggatgttc ggtttattct ttttctttta cttttttatc atgggagcct acttcccgtt
     3721 tttcccgatt tggctacatg acatcaacca tatcagcaaa agtgatacgg gtattatttt
     3781 tgccgctatt tctctgttct cgctattatt ccaaccgctg tttggtctgc tttctgacaa
     3841 actcggaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     3901 ttcacaaata aagcattttt ttcactgcat tctagttgtg gttgtccaaa ctcatcaatg
     3961 tatcttatca tgtctggatc tctgacctga gattggcggc actgaggtag agatgcccga
     4021 acccccccga gggagcgcgg gacgcgccgg ggagggctgg ggccggggag ggctggggcc
     4081 ggggagggct ggggccgggg agggctgggg ccggggaggg ctggggccgg ggagggctgg
     4141 ggccggggag ggctggggct ggggagggct ggggctgggg agggggcggt ggtgtgtagc
     4201 aggagcggtg tgttgcgccg gggtacgtct ggaggagcgg gaggtgcgcg gtgacgtgtg
     4261 gatgaggaac aggagttgtt gcgcggtgag ttgtcgctgt gagttgtgtt gttgggcagg
     4321 tgtggtggat gacgtgacgt gtgacgtgcg gagtgcgccg tgctctgttg gtttcacctg
     4381 tggcagcccg ggccccccgc gggcgcgcgc gcgcgcaaaa aaggcgggcg gcggtccggg
     4441 cggcgtgcgc gcgcgcggcg ggcgtggggg gcggggccgc gggagcgggg ggaggagcgg
     4501 ggggaggagc ggggggagga gcggggggag gagcgggggg aggagcgggg ggaggagcgg
     4561 ggggaggagc ggggggagga gcggggggag gagcgggggg aggagcgggg ggaggagcgg
     4621 ggggaggagc ggggggagga gcggggggag gagcgggggg aggagcgggg ggaggagcgg
     4681 ggggaggagc ggggggagga gcggccagac gccgaaaacg ggcccccccc aaaacacacc
     4741 ccccgggggt cgcgcgcggc cctttaaagc ggtggcggcg ggcagcccgg gccccccgcg
     4801 gccgagacta gcgagttaga caggcaagca ctactcgcct ctgcacgcac atgcttgcct
     4861 gtcaaactct accaccccgg cacgctctct gtctccatgg cccgccgccg ccgccatcgc
     4921 ggcccccgcc gcccccggcc gcccgggccc acgggcgccg tcccaaccgc acagtcccag
     4981 gtaacgagct cgaattaatt cttgaagacg aaagggcctc gtgatacgcc tatttttata
     5041 ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt
     5101 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     5161 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     5221 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     5281 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     5341 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     5401 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg
     5461 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     5521 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     5581 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     5641 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     5701 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg cagcaatggc
     5761 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     5821 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     5881 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     5941 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     6001 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     6061 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     6121 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     6181 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     6241 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     6301 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     6361 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     6421 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     6481 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     6541 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     6601 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     6661 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     6721 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     6781 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaa
//
