pAAVLINKv1-pCALM1-Cre-3'-ROBO1-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-3'-ROBO1-FLAG               7017 bp DNA     circular SYN 19-NOV-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7017)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 2, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..7017
                     /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..3730
                     /codon_start=1
                     /label=ROBO1
                     /translation="VLEQPVGSSSWSSASGFIDTARRETDLLVPTRVSEKSRLLPSHQQ
                     *LTREEAKLSAVEGGLDFSTSVNLPRSHGWQTRGLILATTTMTAPSAAARQAMETATAT
                     SLPTVAQLIV*QIITTNWITNKQI*CSLSQLFMVMWTLVTKSMR*KPSIAQI*RMGVLS
                     IHQGSLLLTPPLSSSSQTSATT*TMAAGTLARSTGNHWDSRNKKWHQFSTTSWSKTS*T
                     KIIEQMTQFLQLSHTTNHTTRTQEDPTTAQTGAVVHLGVRGTRKGQEHPRYQNRVA*TG
                     QTCFLLPQHILLHTAIAKSTTFL*MKAMTKKCHVPCHQQGCICNKMN*KRRKMNEAPLP
                     LFGEQLLLQLPCPIAISPLPL*LPPHRKNSSPCYRIVQRRLATCSTSPTGDGSL*VLLH
                     HHGRSPLHIPMATFQDPWSQIWIRMRQKRKKTKPTWR*PRCKPEGFCYVGLSRHLPPVL
                     GTWRALSRGP*STAGAQPQRRTTFPADAPVLVLRTAPFSLMLTLPRQSQQRQSMLV*K*
                     HDGKCRMLLAVDIFMRLSALGPQVPCLQTAT*VPP*CRKPDQPRN*NTSQDICAEKPTQ
                     MIFHHLLCRHLL*SHLLPNPRHSWKYDL*WCQNSLLWMQEQTDHQTEKEAVTRGEKCWM
                     EDRLLTCEQIQVIPEKHRNSKMTGKDVETRQQNETFHQQRLISSKRIFYLIVDLLFQHQ
                     IIPEIPVPQAQCHQEDQEADKENKQM*VEEILQKCRYLEDMKEEKIIMKN*RKLK"
     misc_feature    1630..1635
                     /label=SDS
     misc_feature    2215..2220
                     /label=SDS
     misc_feature    2336..2341
                     /label=SDS
     misc_feature    2354..2359
                     /label=SDS
     misc_feature    2472..2477
                     /label=SDS
     misc_feature    2669..2674
                     /label=SDS
     misc_feature    2913..2918
                     /label=SDS
     misc_feature    3224..3229
                     /label=SDS
     CDS             3737..3802
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3806..4237
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3950..3955
                     /label=SDS
     repeat_region   4280..4420
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4280..4409
                     /label=AAV2 ITR
     rep_origin      4495..4950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5232..5336
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5337..6197
                     /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      6368..6956
                     /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 ggtattggag cagcctgttg
     1501 gatcatcctc atggtcttca gcatctggct ttatcgacac cgcaagaaga gaaacggact
     1561 tactagtacc tacgcgggta tcagaaaagt cccgtctttt accttcacac caacagtaac
     1621 ttaccagaga ggaggcgaag ctgtcagcag tggagggagg cctggacttc tcaacatcag
     1681 tgaacctgcc gcgcagccat ggctggcaga cacgtggcct aatactggca acaaccacaa
     1741 tgactgctcc atcagctgct gcacggcagg caatggaaac agcgacagca acctcactac
     1801 ctacagtcgc ccagctgatt gtatagcaaa ttataacaac caactggata acaaacaaac
     1861 aaatctgatg ctccctgagt caactgttta tggtgatgtg gaccttagta acaaaatcaa
     1921 tgagatgaaa accttcaata gcccaaatct gaaggatggg cgttttgtca atccatcagg
     1981 gcagcctact ccttacgcca ccactcagct catccagtca aacctcagca acaacatgaa
     2041 caatggcagc ggggactctg gcgagaagca ctggaaacca ctgggacagc agaaacaaga
     2101 agtggcacca gttcagtaca acatcgtgga gcaaaacaag ctgaacaaag attatcgagc
     2161 aaatgacaca gttcctccaa ctatcccata caaccaatca tacgaccaga acacaggagg
     2221 atcctacaac agctcagacc ggggcagtag tacatctggg agtcaggggc acaagaaagg
     2281 ggcaagaaca cccaaggtac caaaacaggg tggcatgaac tgggcagacc tgcttcctcc
     2341 tcccccagca catcctcctc cacacagcaa tagcgaagag tacaacattt ctgtagatga
     2401 aagctatgac caagaaatgc catgtcccgt gccaccagca aggatgtatt tgcaacaaga
     2461 tgaattagaa gaggaggaag atgaacgagg ccccactccc cctgttcggg gagcagcttc
     2521 ttctccagct gccgtgtcct atagccatca gtccactgcc actctgactc cctccccaca
     2581 ggaagaactc cagcccatgt tacaggattg tccagaggag actggccaca tgcagcacca
     2641 gcccgacagg agacggcagc ctgtgagtcc tcctccacca ccacggccga tctcccctcc
     2701 acatacctat ggctacattt caggacccct ggtctcagat atggatacgg atgcgccaga
     2761 agaggaagaa gacgaagccg acatggaggt agccaagatg caaaccagaa ggcttttgtt
     2821 acgtgggctt gagcagacac ctgcctccag tgttggggac ctggagagct ctgtcacggg
     2881 gtccatgatc aacggctggg gctcagcctc agaggaggac aacatttcca gcggacgctc
     2941 cagtgttagt tcttcggacg gctccttttt cactgatgct gactttgccc aggcagtcgc
     3001 agcagcggca gagtatgctg gtctgaaagt agcacgacgg caaatgcagg atgctgctgg
     3061 ccgtcgacat tttcatgcgt ctcagtgccc taggcccaca agtcccgtgt ctacagacag
     3121 caacatgagt gccgccgtaa tgcagaaaac cagaccagcc aagaaactga aacaccagcc
     3181 aggacatctg cgcagagaaa cctacacaga tgatcttcca ccacctcctg tgccgccacc
     3241 tgctataaag tcacctactg cccaatccaa gacacagctg gaagtacgac ctgtagtggt
     3301 gccaaaactc ccttctatgg atgcaagaac agacagatca tcagacagaa aaggaagcag
     3361 ttacaagggg agagaagtgt tggatggaag acaggttgtt gacatgcgaa caaatccagg
     3421 tgatcccaga gaagcacagg aacagcaaaa tgacgggaaa ggacgtggaa acaaggcagc
     3481 aaaacgagac cttccaccag caaagactca tctcatccaa gaggatattc taccttattg
     3541 tagacctact tttccaacat caaataatcc cagagatccc agttcctcaa gctcaatgtc
     3601 atcaagagga tcaggaagca gacaaagaga acaagcaaat gtaggtcgaa gaaatattgc
     3661 agaaatgcag gtacttggag gatatgaaag aggagaagat aataatgaag aattagagga
     3721 aactgaaagc tgtacagact acaaagacca tgacggtgat tataaagatc atgacatcga
     3781 ctacaaggat gacgatgaca agtaactcga gataatcaac ctctggatta caaaatttgt
     3841 gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct
     3901 ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat
     3961 aaatcctggt tagttcttgc cacggcggaa ctcatcgccg cctgccttgc ccgctgctgg
     4021 acaggggctc ggctgttggg cactgacaat tccgtggtgt ttatttgtga aatttgtgat
     4081 gctattgctt tatttgtaac catctagctt tatttgtgaa atttgtgatg ctattgcttt
     4141 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     4201 gtttcaggtt cagggggaga tgtgggaggt tttttaaaga tctggagacc ggtgtggcac
     4261 gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc ctctctgcgc
     4321 gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg
     4381 gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt
     4441 ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca tagtacgcgc
     4501 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac
     4561 ttgccagcgc cttagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg
     4621 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt
     4681 tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc
     4741 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct
     4801 tgttccaaac tggaacaaca ctcaactcta tctcgggcta ttcttttgat ttataaggga
     4861 ttttgccgat ttcggtctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga
     4921 attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca atctgctctg
     4981 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     5041 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     5101 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     5161 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     5221 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     5281 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     5341 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5401 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5461 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5521 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5581 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5641 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5701 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5761 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5821 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5881 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5941 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     6001 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     6061 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     6121 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     6181 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     6241 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     6301 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     6361 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     6421 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     6481 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     6541 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     6601 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     6661 cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc
     6721 gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc
     6781 ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca
     6841 cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc
     6901 tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg
     6961 ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgt
//