LOCUS       sequence-237148-        4996 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of Cre sequence using the CRISPR/Cas9 system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4996)
  AUTHORS   Choi JH, Duboue ER, Macurak M, Chanchu JM, Halpern ME
  TITLE     Specialized neurons in the right habenula mediate response to 
            aversive olfactory cues.
  JOURNAL   Elife. 2021 Dec 8;10:e72345. doi: 10.7554/eLife.72345.
  PUBMED    34878403
REFERENCE   2  (bases 1 to 4996)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2021-12-8"; volume: "10:e72345. doi"; pages: " 
            10.7554/eLife.72345"
COMMENT     Sequence Label: pGEM-T Easy-Gbait-hs-Cre
FEATURES             Location/Qualifiers
     source          1..4996
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      28..37
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      32..41
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      763..772
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             769..1800
                     /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="MANLLTVHQNLPALPVDATSDEVRKNLMDMFRDHQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     polyA_signal    1928..2009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2116..2134)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2117..2134)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2152..2168)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2152..2168)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2165..2187)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2176..2192
                     /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(2200..2206,2207..2224,2225..2230))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2245..2266
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2383..2400)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2554..3142)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2634..2653)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3313..4104,4105..4173))
                     /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     3936..3955
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4174..4278)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4356..4811)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4493..4514)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4705..4724
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4937..4959
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4951..4968
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4952..4968
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4975..4994
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4975..4993
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 aattgggccc gacgtcgcat gctcccggcc gccatggcgg ccgcgggaat tcgatttagg
       61 cgagggcgat gccacctacg gggccgcaat tcactggagg cttccagaac agtgtagagt
      121 ctgaacgggt gcgcgctctg ctgtatttaa agggcgaaag agagaccgca gagaaactca
      181 accgaagaga agcgacttga caaagaagaa aagagcagcc tgacaggact ttttccccga
      241 cgaggtgttt attcgctcta tttaagaatc tactgtaagg taagtctcaa tatattgtac
      301 tctattggct aatcaaaatt atatagagat tatatgtagg ctttaatgtc aaataatcaa
      361 ctttgtatat gtaatctttt tacatgtgga ctgcctataa tgttcatctt attttaggtc
      421 tactagaaaa ttatatttcc cgttttcaca ataaggatta aaaaaacaat gaacagacgg
      481 gcatttactt tatgttgctg acattattat atatgagcat aataaccata aatactagca
      541 aatgtcctaa atgaatttgt gttaatgttg tctacaaaag aaaagaaaat tagcgtttta
      601 cttgtacaac taataataac ttagttattt agagaatttc acttgttgac tagaaaaata
      661 atttaatatt gaaacaattg caccataaat tgtataaata taaaattaat tctaattgtt
      721 ttttttattg cagtttcttg ccgcgggaat tcgattacta gtgccaccat ggccaattta
      781 ctgaccgtac accaaaattt gcctgcatta ccggtcgatg caacgagtga tgaggttcgc
      841 aagaacctga tggacatgtt cagggatcac caggcgtttt ctgagcatac ctggaaaatg
      901 cttctgtccg tttgccggtc gtgggcggca tggtgcaagt tgaataaccg gaaatggttt
      961 cccgcagaac ctgaagatgt tcgcgattat cttctatatc ttcaggcgcg cggtctggca
     1021 gtaaaaacta tccagcaaca tttgggccag ctaaacatgc ttcaccgtcg gtccgggctg
     1081 ccacgaccaa gtgacagcaa tgctgtttca ctggttatgc ggcggatccg aaaagaaaac
     1141 gttgatgccg gtgaacgtgc aaaacaggct ctagcgttcg aacgcactga tttcgaccag
     1201 gttcgttcac tcatggaaaa tagcgatcgc tgccaggata tacgtaatct ggcatttctg
     1261 gggattgctt ataacaccct gttacgtata gccgaaattg ccaggatcag ggttaaagat
     1321 atctcacgta ctgacggtgg gagaatgtta atccatattg gcaggacgaa aacgctggtt
     1381 agcaccgcag gtgtagagaa ggcacttagc ctgggggtaa ctaaactggt cgagcgatgg
     1441 atttccgtct ctggtgtagc tgatgatccg aataactacc tgttttgccg ggtcagaaaa
     1501 aatggtgttg ccgcgccatc tgccaccagc cagctatcaa ctcgcgccct ggaagggatt
     1561 tttgaagcaa ctcatcgatt gatttacggc gctaaggatg actctggtca gagatacctg
     1621 gcctggtctg gacacagtgc ccgtgtcgga gccgcgcgag atatggcccg cgctggagtt
     1681 tcaataccgg agatcatgca agctggtggc tggaccaatg taaatattgt catgaactat
     1741 atccgtaacc tggatagtga aacaggggca atggtgcgcc tgctggaaga tggcgattag
     1801 gctagcgcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     1861 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     1921 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacgaa
     1981 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtccc tgcagaatca
     2041 ctagtgaatt cgcggccgcc tgcaggtcga ccatatggga gagctcccaa cgcgttggat
     2101 gcatagcttg agtattctat agtgtcacct aaatagcttg gcgtaatcat ggtcatagct
     2161 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
     2221 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
     2281 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
     2341 cgcggggaga ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct
     2401 gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt
     2461 atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc
     2521 caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga
     2581 gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata
     2641 ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac
     2701 cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg
     2761 taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc
     2821 cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag
     2881 acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt
     2941 aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt
     3001 atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg
     3061 atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac
     3121 gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca
     3181 gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac
     3241 ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac
     3301 ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     3361 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     3421 accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt
     3481 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     3541 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     3601 tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3661 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3721 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3781 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3841 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3901 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3961 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     4021 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     4081 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     4141 aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat attattgaag
     4201 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     4261 acaaataggg gttccgcgca catttccccg aaaagtgcca cctgatgcgg tgtgaaatac
     4321 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     4381 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     4441 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     4501 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     4561 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     4621 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     4681 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     4741 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     4801 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     4861 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     4921 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga attgtaatac
     4981 gactcactat agggcg
//
