pAAVLINKv1-pCALM1-Cre-NIPBL-C-FLAG

LOCUS       pAAVLINKv1-pCALM1-Cre-NIPBL-C-FLAG                6979 bp DNA     circular SYN 10-AUG-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6979)
  AUTHORS   Stavrini Papadaki
  TITLE     Direct Submission
  JOURNAL   Exported Jun 20, 2026 from SnapGene 6.0.2
            https://www.biofeng.com
FEATURES             Location/Qualifiers
     source          1..6979
                     /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"
                     /note="color: #ffe4c4"
     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..3683
                     /codon_start=1
                     /gene="NIPBL"
                     /gene_synonym="CDLS; CDLS1; IDN3; IDN3-B; Scc2"
                     /product="nipped-B-like protein isoform A"
                     /label=NIPBL
                     /note="isoform A is encoded by transcript variant A;sister 
                     chromatid cohesion 2 homolog; nipped-B-like protein; SCC2 
                     homolog; delangin; Nipped-B homolog"
                     /db_xref="GI:47578105"
                     /db_xref="CCDS:CCDS3920.1"
                     /db_xref="GeneID:25836"
                     /db_xref="HGNC:HGNC:28862"
                     /db_xref="MIM:608667"
                     /protein_id="NP_597677.2"
                     /translation="EDLMKLIIKYGMTVVQHCVSCLGAVVNKVTQNFKFVWACFNRYYG
                     AISKLKSQHQEDPNNTSLLTNKPALLRSLFTVGALCRHFDFDLEDFKGNSKVNIKDKVL
                     ELLMYFTKHSDEEVQTKAIIGLGFAFIQHPSLMFEQEVKNLYNNILSDKNSSVNLKIQV
                     LKNLQTYLQEEDTRMQQADRDWKKVAKQEDLKEMGDVSSGMSSSIMQLYLKQVLEAFFH
                     TQSSVRHFALNVIALTLNQGLIHPVQCVPYLIAMGTDPEPAMRNKADQQLVEIDKKYAG
                     FIHMKAVAGMKMSYQVQQAINTCLKDPVRGFRQDESSSALCSHLYSMIRGNRQHRRAFL
                     ISLLNLFDDTAKTDVTMLLYIADNLACFPYQTQEEPLFIMHHIDITLSVSGSNLLQSFK
                     ESMVKDKRKERKSSPSKENESSDSEEEVSRPRKSRKRVDSDSDSDSEDDINSVMKCLPE
                     NSAPLIEFANVSQGILLLLMLKQHLKNLCGFSDSKIQKYSPSESAKVYDKAINRKTGVH
                     FHPKQTLDFLRSDMANSKITEEVKRSIVKQYLDFKLLMEHLDPDEEEEEGEVSASTNAR
                     NKAITSLLGGGSPKNNTAAETEDDESDGEDRGGGTSGSLRRSKRNSDSTELAAQMNESV
                     DVMDVIAICCPKYKDRPQIARVVQKTSSGFSVQWMAGSYSGSWTEAKRRDGRKLVPWVD
                     TIKESDIIYKKIALTSANKLTNKVVQTLRSLYAAKDGTSS"
     misc_feature    3236..3241
                     /label=SDS
     misc_feature    3302..3307
                     /label=SDS
     misc_feature    3305..3310
                     /label=SDS
     CDS             3687..3752
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      3768..4199
                     /label=W3SL
                     /note="Short WPRE and SV40 polyA"
     misc_feature    3912..3917
                     /label=SDS
     repeat_region   4242..4382
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     misc_feature    4242..4371
                     /label=AAV2 ITR
     repeat_region   4243..4383
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      4457..4912
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5194..5298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5299..6159
                     /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      6330..6918
                     /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 ggaagatcta atgaagctca
     1501 tcatcaaata tggcatgact gtagtgcaac attgtgtgag ctgtcttgga gctgttgtaa
     1561 ataaagtgac acaaaatttt aaatttgtgt gggcttgttt caatagatac tatggtgcca
     1621 tttcaaaatt aaaaagtcaa caccaagagg acccaaataa cacttcactt ctaacaaaca
     1681 aaccagcact tcttagatcc cttttcaccg ttggagcact atgtcggcat tttgattttg
     1741 atctggaaga ttttaaaggc aacagcaagg ttaacataaa agataaagta cttgaactat
     1801 tgatgtattt tacaaaacac tcagatgaag aagtacaaac aaaagctatc attggtctag
     1861 gatttgcctt tattcagcat ccaagtctaa tgttcgagca agaagtgaag aatctatata
     1921 ataatatttt atctgataag aactcctcag tcaatttaaa aatacaagtg ttaaaaaacc
     1981 tccagaccta cctacaagaa gaagatacac gtatgcagca ggcagataga gactggaaga
     2041 aagttgcaaa acaggaagac ttaaaagaaa tgggtgatgt ttcctcaggg atgagtagtt
     2101 ccatcatgca gctttatctc aaacaggtgc ttgaggcatt ttttcacacc cagtcaagtg
     2161 tacgccactt tgccctaaat gtcattgcat tgactctaaa tcaaggtctt attcatccag
     2221 ttcagtgtgt gccatattta attgctatgg gcacagaccc agaacctgct atgcggaaca
     2281 aggctgatca gcaacttgtg gaaatagaca aaaaatatgc tggattcatt catatgaaag
     2341 cagtggctgg tatgaagatg tcttaccagg tacaacaggc aatcaacaca tgcctaaaag
     2401 atcctgtaag gggtttcaga caagacgagt cctctagcgc tttgtgttca cacctttact
     2461 ccatgatccg tggaaaccgc caacacagac gagcctttct tatttcttta ctcaacctct
     2521 ttgatgacac agcaaaaaca gacgtgacta tgctcttgta tatagcagac aatctagcct
     2581 gttttccata ccagacacag gaagagccgt tgtttataat gcatcatata gacattacac
     2641 tctcagtttc tggtagtaac ctactgcagt cattcaagga gtctatggta aaggacaaaa
     2701 ggaaagagag aaaatcatca cctagtaagg aaaatgagtc aagcgacagt gaagaagaag
     2761 tttccaggcc tcggaagtca cggaaacgtg tagattcaga ttcagattca gattcagaag
     2821 acgatataaa ttcagtgatg aaatgtttgc cagaaaattc agctccttta atcgaatttg
     2881 caaatgtgtc ccagggtatt ttattacttc tcatgttaaa acaacatttg aagaatcttt
     2941 gtggattttc tgatagtaaa attcagaagt actctccatc tgaatctgca aaagtatatg
     3001 ataaagcgat aaaccgaaaa acaggagttc attttcatcc aaaacaaaca ctggacttcc
     3061 tgcggagtga catggctaat tccaaaatca cagaagaggt gaaaaggagt atagtaaaac
     3121 agtatctaga tttcaaactt ctcatggaac atctggaccc tgatgaagaa gaagaagaag
     3181 gggaggtttc agctagcaca aatgctcgga acaaagcaat tacctcactg cttggaggag
     3241 gcagccctaa aaataataca gcagcagaga cagaagatga tgaaagtgat ggggaggata
     3301 gaggaggagg cacttcaggg tcattgagaa ggtcaaaacg aaattcagac tctacggagt
     3361 tggcagcaca gatgaatgaa agtgttgacg tcatggatgt catcgctatt tgctgtccaa
     3421 agtacaaaga tcgaccacaa attgcaagag tagtgcagaa aaccagcagt ggcttcagtg
     3481 ttcagtggat ggcaggctcc tacagtggct cctggactga ggctaagcgc cgtgatggcc
     3541 gcaaactggt gccttgggta gacactatta aagagtcaga cattatttac aaaaaaattg
     3601 ctctaacgag tgctaataag ctgactaata aagttgttca gactttacga tccctgtatg
     3661 ccgccaagga tgggacttcc agcaaggact acaaagacca tgacggtgat tataaagatc
     3721 atgacatcga ctacaaggat gacgatgaca agtaagaatt ctgatgactc gagataatca
     3781 acctctggat tacaaaattt gtgaaagatt gactggtatt cttaactatg ttgctccttt
     3841 tacgctatgt ggatacgctg ctttaatgcc tttgtatcat gctattgctt cccgtatggc
     3901 tttcattttc tcctccttgt ataaatcctg gttagttctt gccacggcgg aactcatcgc
     3961 cgcctgcctt gcccgctgct ggacaggggc tcggctgttg ggcactgaca attccgtggt
     4021 gtttatttgt gaaatttgtg atgctattgc tttatttgta accatctagc tttatttgtg
     4081 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttaaca
     4141 acaacaattg cattcatttt atgtttcagg ttcaggggga gatgtgggag gttttttaaa
     4201 gatctggaga ccggtgtggc acgtgcggac cgagcggccg caggaacccc tagtgatgga
     4261 gttggccact ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc
     4321 ccgacgcccg ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gctgcctgca
     4381 ggggcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatacg
     4441 tcaaagcaac catagtacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     4501 acgcgcagcg tgaccgctac acttgccagc gccttagcgc ccgctccttt cgctttcttc
     4561 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     4621 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga tttgggtgat
     4681 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     4741 acgttcttta atagtggact cttgttccaa actggaacaa cactcaactc tatctcgggc
     4801 tattcttttg atttataagg gattttgccg atttcggtct attggttaaa aaatgagctg
     4861 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgtttacaat tttatggtgc
     4921 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4981 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5041 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5101 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5161 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5221 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5281 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5341 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5401 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5461 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5521 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5581 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5641 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5701 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5761 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5821 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5881 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5941 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6001 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6061 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6121 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6181 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6241 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6781 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6841 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6901 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6961 tggccttttg ctcacatgt
//