LOCUS       sequence-233842-        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      33..42
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      37..46
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      768..777
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             774..1805
                     /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
                     RWISVSGVADDPNNYLFCRVRKXGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     polyA_signal    1933..2014
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     promoter        complement(2121..2139)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2122..2139)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2157..2173)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2157..2173)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2170..2192)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2181..2197
                     /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(2205..2211,2212..2229,2230..2235))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2250..2271
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2388..2405)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2559..3147)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2639..2658)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3318..4109,4110..4178))
                     /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     3941..3960
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4179..4283)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4361..4816)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4498..4519)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4710..4729
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4942..4964
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     4956..4973
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4957..4973
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4980..4996,1..3)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        join(4980..4996,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 gggcgaattg ggcccgacgt cgcatgctcc cggccgccat ggcggccgcg ggaattcgat
       61 ttaggcgagg gcgatgccac ctacggggcc gcaattcact ggaggcttcc agaacagtgt
      121 agagtctgaa cgggtgcgcg ctctgctgta tttaaagggc gaaagagaga ccgcagagaa
      181 actcaaccga agagaagcga cttgacaaag aagaaaagag cagcctgaca ggactttttc
      241 cccgacgagg tgtttattcg ctctatttaa gaatctactg taaggtaagt ctcaatatat
      301 tgtactctat tggctaatca aaattatata gagattatat gtaggcttta atgtcaaata
      361 atcaactttg tatatgtaat ctttttacat gtggactgcc tataatgttc atcttatttt
      421 aggtctacta gaaaattata tttcccgttt tcacaataag gattaaaaaa acaatgaaca
      481 gacgggcatt tactttatgt tgctgacatt attatatatg agcataataa ccataaatac
      541 tagcaaatgt cctaaatgaa tttgtgttaa tgttgtctac aaaagaaaag aaaattagcg
      601 ttttacttgt acaactaata ataacttagt tatttagaga atttcacttg ttgactagaa
      661 aaataattta atattgaaac aattgcacca taaattgtat aaatataaaa ttaattctaa
      721 ttgttttttt tattgcagtt tcttgccgcg ggaattcgat tactagtgcc accatggcca
      781 atttactgac cgtacaccaa aatttgcctg cattaccggt cgatgcaacg agtgatgagg
      841 ttcgcaagaa cctgatggac atgttcaggg atcaccaggc gttttctgag catacctgga
      901 aaatgcttct gtccgtttgc cggtcgtggg cggcatggtg caagttgaat aaccggaaat
      961 ggtttcccgc agaacctgaa gatgttcgcg attatcttct atatcttcag gcgcgcggtc
     1021 tggcagtaaa aactatccag caacatttgg gccagctaaa catgcttcat cgtcggtccg
     1081 ggctgccacg accaagtgac agcaatgctg tttcactggt tatgcggcgg atccgaaaag
     1141 aaaacgttga tgccggtgaa cgtgcaaaac aggctctagc gttcgaacgc actgatttcg
     1201 accaggttcg ttcactcatg gaaaatagcg atcgctgcca ggatatacgt aatctggcat
     1261 ttctggggat tgcttataac accctgttac gtatagccga aattgccagg atcagggtta
     1321 aagatatctc acgtactgac ggtgggagaa tgttaatcca tattggcaga acgaaaacgc
     1381 tggttagcac cgcaggtgta gagaaggcac ttagcctggg ggtaactaaa ctggtcgagc
     1441 gatggatttc cgtctctggt gtagctgatg atccgaataa ctacctgttt tgccgggtca
     1501 gaaaaantgg tgttgccgcg ccatctgcca ccagccagct atcaactcgc gccctggaag
     1561 ggatttttga agcaactcat cgattgattt acggcgctaa ggatgactct ggtcagagat
     1621 acctggcctg gtctggacac agtgcccgtg tcggagccgc gcgagatatg gcccgcgctg
     1681 gagtttcaat accggagatc atgcaagctg gtggctggac caatgtaaat attgtcatga
     1741 actatatccg taacctggat agtgaaacag gggcaatggt gcgcctgctg gaagatggcg
     1801 attaggctag cgcggccgcg actctagatc ataatcagcc ataccacatt tgtagaggtt
     1861 ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa aatgaatgca
     1921 attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     1981 acgaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccctgcag
     2041 aatcactagt gaattcgcgg ccgcctgcag gtcgaccata tgggagagct cccaacgcgt
     2101 tggatgcata gcttgagtat tctatagtgt cacctaaata gcttggcgta atcatggtca
     2161 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2221 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2281 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2341 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2401 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2461 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2521 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2581 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2641 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2701 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     2761 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     2821 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2881 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     2941 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3001 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3061 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3121 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3181 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3241 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3301 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3361 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     3421 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     3481 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     3541 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     3601 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     3661 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     3721 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     3781 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     3841 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     3901 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     3961 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     4021 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     4081 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     4141 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     4201 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     4261 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga tgcggtgtga
     4321 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag cgttaatatt
     4381 ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa
     4441 atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca
     4501 gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc
     4561 gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg
     4621 aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg
     4681 ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg
     4741 gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg
     4801 ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg
     4861 tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca aggcgattaa
     4921 gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc agtgaattgt
     4981 aatacgactc actata
//
