pAAVLINKv1-pCALM1-Cre-3'-SCN4A-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-SCN4A-FLAG               7006 bp DNA     circular SYN 17-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7006)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported May 18, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7006
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    12..130
                     /label=AAV2 ITR
     promoter        145..265
                     /label=pCALM1
     CDS             277..1308
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     intron          1309..1349
                     /label=Splicing Donor
     protein_bind    1356..1389
                     /label=lox2272-JTZ17
                     /bound_moiety="Cre recombinase"
                     /note="Right element (RE) mutant of loxP (Araki et al., 
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     intron          1390..1481
                     /label=Splicing Acceptor
                     /note="Splicing acceptor"
     CDS             1482..3719
                     /codon_start=1
                     /label=SCN4A
                     /translation="VYFTNAWCWLDFLIVDVSIISLVANWLGYSELGPIKSLRTLRALR
                     PLRALSRFEGMRVVVNALLGAIPSIMNVLLVCLIFWLIFSIMGVNLFAGKFYYCINTTT
                     SERFDISEVNNKSECESLMHTGQVRWLNVKVNYDNVGLGYLSLLQVATFKGWMDIMYAA
                     VDSREKEEQPQYEVNLYMYLYFVIFIIFGSFFTLNLFIGVIIDNFNQQKKKLGGKDIFM
                     TEEQKKYYNAMKKLGSKKPQKPIPRPQNKIQGMVYDLVTKQAFDITIMILICLNMVTMM
                     VETDNQSQLKVDILYNINMIFIIIFTGECVLKMLALRQYYFTVGWNIFDFVVVILSIVG
                     LALSDLIQKYFVSPTLFRVIRLARIGRVLRLIRGAKGIRTLLFALMMSLPALFNIGLLL
                     FLVMFIYSIFGMSNFAYVKKESGIDDMFNFETFGNSIICLFEITTSAGWDGLLNPILNS
                     GPPDCDPNLENPGTSVKGDCGNPSIGICFFCSYIIISFLIVVNMYIAIILENFNVATEE
                     SSEPLGEDDFEMFYETWEKFDPDATQFIAYSRLSDFVDTLQEPLRIAKPNKIKLITLDL
                     PMVPGDKIHCLDILFALTKEVLGDSGEMDALKQTMEEKFMAANPSKVSYEPITTTLKRK
                     HEEVCAIKIQRAYRRHLLQRSMKQASYMYRHSHDGSGDDAPEKEGLLANTMSKMYGHEN
                     GNSSSPSPEEKGEAGDAGPTMGLMPISPSDTAWPPAPPPGQTVRPGVKESLV"
     misc_feature    1667..1672
                     /label=SDS
     misc_feature    1919..1924
                     /label=SDS
     misc_feature    1987..1992
                     /label=SDS
     misc_feature    2669..2674
                     /label=SDS
     misc_feature    2672..2677
                     /label=SDS
     misc_feature    3391..3396
                     /label=SDS
     misc_feature    3514..3519
                     /label=SDS
     CDS             3726..3791
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3795..4226
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3939..3944
                     /label=SDS
     repeat_region   4269..4409
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4269..4398
                     /label=AAV2 ITR
     rep_origin      4484..4939
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5221..5325
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5326..6186
                     /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"
     rep_origin      6357..6945
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtcgctct gcgcgaggcg gctccgccgc ggcggaggga
      181 tacggcgcac catatatata tcgcggggcg cagactcgcg ctccggcagt ggtgctggga
      241 gtgtcgtgga cgccgtgccg ttactcgtac gccaccatgt ccaatttact gaccgtacac
      301 caaaatttgc ctgcattacc ggtcgatgca acgagtgatg aggttcgcaa gaacctgatg
      361 gacatgttca gggatcgcca ggcgttttct gagcatacct ggaaaatgct tctgtccgtt
      421 tgccggtcgt gggcggcatg gtgcaagttg aataaccgga aatggtttcc cgcagaacct
      481 gaagatgttc gcgattatct tctatatctt caggcgcgcg gtctggcagt aaaaaccatc
      541 cagcaacatt tgggccagct aaacatgctt catcgtcggt ccgggctgcc acgaccaagt
      601 gacagcaatg ctgtttcact ggttatgcgg cggatccgaa aagaaaacgt tgatgccggt
      661 gaacgtgcaa aacaggctct agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc
      721 atggaaaata gcgatcgctg ccaggatata cgtaatctgg catttctggg gattgcttat
      781 aacaccctgt tacgtatagc cgaaattgcc aggatcaggg ttaaagatat ctcacgtact
      841 gacggtggga gaatgttaat ccatattggc agaacgaaaa cgctggttag caccgcaggt
      901 gtagagaagg cacttagcct gggggtaact aaactggtcg agcgatggat ttccgtctct
      961 ggtgtagctg atgatccgaa taactacctg ttttgccggg tcagaaaaaa tggtgttgcc
     1021 gcgccatctg ccaccagcca gctatcaact cgcgccctgg aagggatttt tgaagcaact
     1081 catcgattga tttacggcgc taaggatgac tctggtcaga gatacctggc ctggtctgga
     1141 cacagtgccc gtgtcggagc cgcgcgagat atggcccgcg ctggagtttc aataccggag
     1201 atcatgcaag ctggtggctg gaccaatgta aatattgtaa tgaactatat ccgtaacctg
     1261 gatagtgaaa caggggcaat ggtgcgcctg ctggaagatg gcgattaggt aagtatcaag
     1321 gttacaagac aggtttaagg agaccaataa ctagtataac ttcgtatagg atactttata
     1381 gcaatttatg aaactgggct tgtcgagaca gagaagactc ttgcgtttct gataggcacc
     1441 tattggtctt actgacatcc actttgcctt tctctccaca ggtgtacttc accaacgcct
     1501 ggtgctggct cgacttcctc atcgtggatg tctccatcat cagcttggtg gccaactggc
     1561 tgggctactc ggagctggga cccatcaaat ccctgcggac actgcgggcc ctgcgtcccc
     1621 tgagggcact gtcccgattc gagggcatga gggtggtggt gaacgccctc ctaggcgcca
     1681 tcccctccat catgaatgtg ctgcttgtct gcctcatctt ctggctgatc ttcagcatca
     1741 tgggtgtcaa cctgtttgcc ggcaagttct actactgcat caacaccacc acctctgaga
     1801 ggttcgacat ctccgaggtc aacaacaagt ctgagtgcga gagcctcatg cacacaggcc
     1861 aggtccgctg gctcaatgtc aaggtcaact acgacaacgt gggtctgggc tacctctccc
     1921 tcctgcaggt ggccaccttc aagggttgga tggacatcat gtatgcagcc gtggactccc
     1981 gggagaagga ggagcagccg cagtacgagg tgaacctcta catgtacctc tactttgtca
     2041 tcttcatcat ctttggctcc ttcttcaccc tcaacctctt cattggcgtc atcattgaca
     2101 acttcaacca gcagaagaag aagttagggg ggaaagacat ctttatgacg gaggaacaga
     2161 agaaatacta taacgccatg aagaagcttg gctccaagaa gcctcagaag ccaattcccc
     2221 ggccccagaa caagatccag ggcatggtgt atgacctcgt gacgaagcag gccttcgaca
     2281 tcaccatcat gatcctcatc tgcctcaaca tggtcaccat gatggtggag acagacaacc
     2341 agagccagct caaggtggac atcctgtaca acatcaacat gatcttcatc atcatcttca
     2401 caggggagtg cgtgctcaag atgctcgccc tgcgccagta ctacttcacc gttggctgga
     2461 acatctttga cttcgtggtc gtcatcctgt ccattgtggg ccttgccctc tctgacctga
     2521 tccagaagta cttcgtgtca cccacgctgt tccgtgtgat ccgcctggcg cggattgggc
     2581 gtgtcctgcg gctgatccgc ggggccaagg gcatccggac gctgctgttc gccctcatga
     2641 tgtcgctgcc tgccctcttc aacatcggcc tcctcctctt cctggtcatg ttcatctact
     2701 ccatcttcgg catgtccaac tttgcctacg tcaagaagga gtcgggcatc gatgatatgt
     2761 tcaacttcga gaccttcggc aacagcatca tctgcctgtt cgagatcacc acgtcggccg
     2821 gctgggacgg gctcctcaac cccatcctca acagcgggcc cccagactgt gaccccaacc
     2881 tggagaaccc gggcaccagt gtcaagggtg actgcggcaa cccctccatc ggcatctgct
     2941 tcttctgcag ctatatcatc atctccttcc tcatcgtggt caacatgtac atcgccatca
     3001 tcctggagaa cttcaatgtg gccacagagg agagcagcga gccccttggt gaagatgact
     3061 ttgagatgtt ctacgagaca tgggagaagt tcgaccccga cgccacccag ttcatcgcct
     3121 acagccgcct ctcagacttc gtggacaccc tgcaggaacc gctgaggatt gccaagccca
     3181 acaagatcaa gctcatcaca ctggacttgc ccatggtgcc aggggacaag atccactgcc
     3241 tggacatcct ctttgccctg accaaagagg tcctgggtga ctctggggaa atggacgccc
     3301 tcaagcagac catggaggag aagttcatgg cagccaaccc ctccaaggtg tcctacgagc
     3361 ccatcaccac caccctcaag aggaagcacg aggaggtgtg cgccatcaag atccagaggg
     3421 cctaccgccg gcacctgcta cagcgctcca tgaagcaggc atcctacatg taccgccaca
     3481 gccacgacgg cagcggggat gacgcccctg agaaggaggg gctgcttgcc aacaccatga
     3541 gcaagatgta tggccacgag aatgggaaca gcagctcgcc aagcccggag gagaagggcg
     3601 aggcagggga cgccggaccc actatggggc tgatgcccat cagcccctca gacactgcct
     3661 ggcctcccgc ccctccccca gggcagactg tgcgcccagg tgtcaaggag tctcttgtct
     3721 gtacagacta caaagaccat gacggtgatt ataaagatca tgacatcgac tacaaggatg
     3781 acgatgacaa gtaactcgag ataatcaacc tctggattac aaaatttgtg aaagattgac
     3841 tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt
     3901 gtatcatgct attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt
     3961 agttcttgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga caggggctcg
     4021 gctgttgggc actgacaatt ccgtggtgtt tatttgtgaa atttgtgatg ctattgcttt
     4081 atttgtaacc atctagcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     4141 ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc
     4201 agggggagat gtgggaggtt ttttaaagat ctggagaccg gtgtggcacg tgcggaccga
     4261 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     4321 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     4381 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     4441 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     4501 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4561 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4621 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4681 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4741 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4801 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     4861 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     4921 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     4981 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5041 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5101 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     5161 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     5221 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     5281 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     5341 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     5401 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     5461 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     5521 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     5581 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     5641 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     5701 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     5761 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     5821 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     5881 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     5941 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6001 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6061 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6121 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     6181 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     6241 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     6301 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     6361 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     6421 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     6481 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     6541 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     6601 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     6661 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     6721 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     6781 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     6841 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     6901 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     6961 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgt
//