LOCUS       sequence-319332-        8291 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses IntCCCre in neurons in split intein-mediated split-Cre 
            recombinase applications.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8291)
  AUTHORS   Khoo ATT, Kim PJ, Kim HM, Je HS
  TITLE     Neural circuit analysis using a novel intersectional split 
            intein-mediated split-Cre recombinase system.
  JOURNAL   Mol Brain. 2020 Jul 2;13(1):101. doi: 10.1186/s13041-020-00640-2.
  PUBMED    32616061
REFERENCE   2  (bases 1 to 8291)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol 
            Brain."; date: "2020-07-2"; volume: "13(1):101. doi"; pages: " 
            10.1186/s13041-020-00640-2"
COMMENT     Sequence Label: pCDH-hSyn-IntCCCre-T2A-CopGFP
FEATURES             Location/Qualifiers
     source          1..8291
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     173..191
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(229..251)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     351..370
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     564..586
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     578..595
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     579..595
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        610..836
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             837..1017
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1064..1189
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1682..1915
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     CDS             2100..2144
                     /codon_start=1
                     /product="antigenic peptide corresponding to amino acids 
                     655 to 669 of the HIV envelope protein gp41 (Lutje Hulsik 
                     et al., 2013)"
                     /label=gp41 peptide
                     /note="recognized by the 2H10 single-chain llama nanobody"
                     /translation="KNEQELLELDKWASL"
     misc_feature    2411..2527
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     promoter        2566..3013
                     /label=hSyn promoter
                     /note="human synapsin I promoter; confers neuron-specific 
                     expression (Kugler et al., 2003)"
     regulatory      3029..3038
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3038..3061
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4088..4111
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             4121..4174
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid 
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4205..4867
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
     misc_feature    4974..5562
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5027..5047)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(5445..5456)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     LTR             5636..5869
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    5941..6062
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5978..5997)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6032..6051
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      6102..6237
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6270..6286)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6270..6286)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6283..6305)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6294..6310
                     /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(6318..6324,6325..6342,6343..6348))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6363..6384
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6501..6518)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6672..7260)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(7431..8222,8223..8291))
                     /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     8054..8073
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata aaaataggcg
      181 tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
      241 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
      301 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
      361 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
      421 aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc
      481 ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt
      541 aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgccaa
      601 gctgacgcgt gtagtcttat gcaatactct tgtagtcttg caacatggta acgatgagtt
      661 agcaacatgc cttacaagga gagaaaaagc accgtgcatg ccgattggtg gaagtaaggt
      721 ggtacgatcg tgccttatta ggaaggcaac agacgggtct gacatggatt ggacgaacca
      781 ctgaattgcc gcattgcaga gatattgtat ttaagtgcct agctcgatac ataaacgggt
      841 ctctctggtt agaccagatc tgagcctggg agctctctgg ctaactaggg aacccactgc
      901 ttaagcctca ataaagcttg ccttgagtgc ttcaagtagt gtgtgcccgt ctgttgtgtg
      961 actctggtaa ctagagatcc ctcagaccct tttagtcagt gtggaaaatc tctagcagtg
     1021 gcgcccgaac agggacttga aagcgaaagg gaaaccagag gagctctctc gacgcaggac
     1081 tcggcttgct gaagcgcgca cggcaagagg cgaggggcgg cgactggtga gtacgccaaa
     1141 aattttgact agcggaggct agaaggagag agatgggtgc gagagcgtca gtattaagcg
     1201 ggggagaatt agatcgcgat gggaaaaaat tcggttaagg ccagggggaa agaaaaaata
     1261 taaattaaaa catatagtat gggcaagcag ggagctagaa cgattcgcag ttaatcctgg
     1321 cctgttagaa acatcagaag gctgtagaca aatactggga cagctacaac catcccttca
     1381 gacaggatca gaagaactta gatcattata taatacagta gcaaccctct attgtgtgca
     1441 tcaaaggata gagataaaag acaccaagga agctttagac aagatagagg aagagcaaaa
     1501 caaaagtaag accaccgcac agcaagcggc cactgatctt cagacctgga ggaggagata
     1561 tgagggacaa ttggagaagt gaattatata aatataaagt agtaaaaatt gaaccattag
     1621 gagtagcacc caccaaggca aagagaagag tggtgcagag agaaaaaaga gcagtgggaa
     1681 taggagcttt gttccttggg ttcttgggag cagcaggaag cactatgggc gcagcgtcaa
     1741 tgacgctgac ggtacaggcc agacaattat tgtctggtat agtgcagcag cagaacaatt
     1801 tgctgagggc tattgaggcg caacagcatc tgttgcaact cacagtctgg ggcatcaagc
     1861 agctccaggc aagaatcctg gctgtggaaa gatacctaaa ggatcaacag ctcctgggga
     1921 tttggggttg ctctggaaaa ctcatttgca ccactgctgt gccttggaat gctagttgga
     1981 gtaataaatc tctggaacag atttggaatc acacgacctg gatggagtgg gacagagaaa
     2041 ttaacaatta cacaagctta atacactcct taattgaaga atcgcaaaac cagcaagaaa
     2101 agaatgaaca agaattattg gaattagata aatgggcaag tttgtggaat tggtttaaca
     2161 taacaaattg gctgtggtat ataaaattat tcataatgat agtaggaggc ttggtaggtt
     2221 taagaatagt ttttgctgta ctttctatag tgaatagagt taggcaggga tattcaccat
     2281 tatcgtttca gacccacctc ccaaccccga ggggacccga caggcccgaa ggaatagaag
     2341 aagaaggtgg agagagagac agagacagat ccattcgatt agtgaacgga tctcgacggt
     2401 atcggttaac ttttaaaaga aaagggggga ttggggggta cagtgcaggg gaaagaatag
     2461 tagacataat agcaacagac atacaaacta aagaattaca aaaacaaatt acaaaattca
     2521 aaattttatc gatatcgatc tagactgcag agggccctgc gtatgagtgc aagtgggttt
     2581 taggaccagg atgaggcggg gtgggggtgc ctacctgacg accgaccccg acccactgga
     2641 caagcaccca acccccattc cccaaattgc gcatccccta tcagagaggg ggaggggaaa
     2701 caggatgcgg cgaggcgcgt gcgcactgcc agcttcagca ccgcggacag tgccttcgcc
     2761 cccgcctggc ggcgcgcgcc accgccgcct cagcactgaa ggcgcgctga cgtcactcgc
     2821 cggtcccccg caaactcccc ttcccggcca ccttggtcgc gtccgcgccg ccgccggccc
     2881 agccggaccg caccacgcga ggcgcgagat aggggggcac gggcgcgacc atctgcgctg
     2941 cggcgccggc gactcagcgc tgcctcagtc tgcggtgggc agcggaggag tcgtgtcgtg
     3001 cctgagagcg cagtcgagaa ttaattaagc caccatggat tataaagacg atgacgataa
     3061 aatggttaaa gttatcggtc gtcgttccct cggagtgcaa agaatatttg atattggtct
     3121 tcgccaagac cataattttc tgctagccaa tggggcgatc gcggccaatt gcttcaataa
     3181 ggaggacgga tccaaccgga aatggtttcc cgcagaacct gaagatgttc gcgattatct
     3241 tctatatctt caggcgcgcg gtctggcagt aaaaactatc cagcaacatt tgggccagct
     3301 aaacatgctt catcgtcggt ccgggctgcc acgaccaagt gacagcaatg ctgtttcact
     3361 ggttatgcgg cggatacgaa aagaaaacgt tgatgccggt gaacgtgcaa aacaggctct
     3421 agcgttcgaa cgcactgatt tcgaccaggt tcgttcactc atggaaaata gcgatcgctg
     3481 ccaggatata cgtaatctgg catttctggg gattgcttat aacaccctgt tacgtatagc
     3541 cgaaattgcc aggatcaggg ttaaagatat ctcacgtact gacggtggga gaatgttaat
     3601 ccatattggc agaacgaaaa cgctggttag caccgcaggt gtagagaagg cacttagcct
     3661 gggggtaact aaactggtcg agcgatggat ttccgtctct ggtgtagctg atgatccgaa
     3721 taactacctg ttttgccggg tcagaaaaaa tggtgttgcc gcgccatctg ccaccagcca
     3781 gctatcaact cgcgccctgg aagggatttt tgaagcaact catcgattga tttacggcgc
     3841 taaggatgac tctggtcaga gatacctggc ctggtctgga cacagtgccc gtgtcggagc
     3901 cgcgcgagat atggcccgcg ctggagtttc aataccggag atcatgcaag ctggtggctg
     3961 gaccaatgta aatattgtca tgaactatat ccgtaacctg gatagtgaaa caggggcaat
     4021 ggtgcgcctg ctggaagatg gcgatgaatt ctgcagatat ccatcacact ggcggccgct
     4081 cgagatggac tacaaggacg acgatgataa gggcaccggt gagggcagag gaagtcttct
     4141 aacatgcggt gacgtggagg agaatcccgg ccctcccggg atggagagcg acgagagcgg
     4201 cctgcccgcc atggagatcg agtgccgcat caccggcacc ctgaacggcg tggagttcga
     4261 gctggtgggc ggcggagagg gcacccccaa gcagggccgc atgaccaaca agatgaagag
     4321 caccaaaggc gccctgacct tcagccccta cctgctgagc cacgtgatgg gctacggctt
     4381 ctaccacttc ggcacctacc ccagcggcta cgagaacccc ttcctgcacg ccatcaacaa
     4441 cggcggctac accaacaccc gcatcgagaa gtacgaggac ggcggcgtgc tgcacgtgag
     4501 cttcagctac cgctacgagg ccggccgcgt gatcggcgac ttcaaggtgg tgggcaccgg
     4561 cttccccgag gacagcgtga tcttcaccga caagatcatc cgcagcaacg ccaccgtgga
     4621 gcacctgcac cccatgggcg ataacgtgct ggtgggcagc ttcgcccgca ccttcagcct
     4681 gcgcgacggc ggctactaca gcttcgtggt ggacagccac atgcacttca agagcgccat
     4741 ccaccccagc atcctgcaga acgggggccc catgttcgcc ttccgccgcg tggaggagct
     4801 gcacagcaac accgagctgg gcatcgtgga gtaccagcac gccttcaaga cccccatcgc
     4861 cttcgccaga tcccgcgctc agtcgtccaa ttctgccgtg gacggcaccg ccggacccgg
     4921 ctccaccctg tacaagtaac acctggtgca tgacccgcaa gcccggtgcc tgaaatcaac
     4981 ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta
     5041 cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt
     5101 tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg
     5161 ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg
     5221 gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca
     5281 cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca
     5341 ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg
     5401 ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag
     5461 cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt ctccgccttc
     5521 gccctcagac gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca
     5581 atgacttaca aggcagctgt agatcttagc cactttttaa aagaaaaggg gggactggaa
     5641 gggctaattc actcccaacg aagataagat ctgctttttg cttgtactgg gtctctctgg
     5701 ttagaccaga tctgagcctg ggagctctct ggctaactag ggaacccact gcttaagcct
     5761 caataaagct tgccttgagt gcttcaagta gtgtgtgccc gtctgttgtg tgactctggt
     5821 aactagagat ccctcagacc cttttagtca gtgtggaaaa tctctagcag tagtagttca
     5881 tgtcatctta ttattcagta tttataactt gcaaagaaat gaatatcaga gagtgagagg
     5941 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     6001 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     6061 tatcatgtct ggctctagct atcccgcccc taactccgcc catcccgccc ctaactccgc
     6121 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6181 aggccgcctc ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6241 acttttgcag agaccaaatt cgtaatcatg tcatagctgt ttcctgtgtg aaattgttat
     6301 ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc
     6361 taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     6421 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     6481 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     6541 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     6601 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     6661 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     6721 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     6781 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     6841 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     6901 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     6961 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     7021 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     7081 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     7141 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     7201 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     7261 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     7321 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     7381 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     7441 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     7501 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     7561 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     7621 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     7681 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     7741 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     7801 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     7861 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     7921 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     7981 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     8041 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     8101 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     8161 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     8221 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     8281 tgaatactca t
//
