LOCUS       sequence-237152-        5057 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Integration of QF2 sequence using the CRISPR/Cas9 system..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5057)
  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 5057)
  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-QF2
FEATURES             Location/Qualifiers
     source          1..5057
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     6..23
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7..23
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     30..49
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        30..48
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      79..88
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      83..92
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     protein_bind    complement(1898..1922)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     polyA_signal    1924..2058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1948..1967)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2002..2021
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     protein_bind    complement(2121..2141)
                     /gene="mutant version of attB"
                     /label=attB3
                     /bound_moiety="BP Clonase(TM)"
                     /note="core recombination site for the Gateway(R) BP 
                     reaction"
     promoter        complement(2228..2246)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2229..2246)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2264..2280)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2264..2280)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2277..2299)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2288..2304
                     /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(2312..2318,2319..2336,2337..2342))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2357..2378
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2495..2512)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2666..3254)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2746..2765)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3425..4216,4217..4285))
                     /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     4048..4067
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4286..4390)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4468..4923)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4605..4626)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4817..4836
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     join(5049..5057,1..14)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc gaattgggcc
       61 cgacgtcgca tgctcccggc cgccatggcg gccgcgggaa ttcgatttag gcgagggcga
      121 tgccacctac ggggccgcaa ttcactggag gcttccagaa cagtgtagag tctgaacggg
      181 tgcgcgctct gctgtattta aagggcgaaa gagagaccgc agagaaactc aaccgaagag
      241 aagcgacttg acaaagaaga aaagagcagc ctgacaggac tttttccccg acgaggtgtt
      301 tattcgctct atttaagaat ctactgtaag gtaagtctca atatattgta ctctattggc
      361 taatcaaaat tatatagaga ttatatgtag gctttaatgt caaataatca actttgtata
      421 tgtaatcttt ttacatgtgg actgcctata atgttcatct tattttaggt ctactagaaa
      481 attatatttc ccgttttcac aataaggatt aaaaaaacaa tgaacagacg ggcatttact
      541 ttatgttgct gacattatta tatatgagca taataaccat aaatactagc aaatgtccta
      601 aatgaatttg tgttaatgtt gtctacaaaa gaaaagaaaa ttagcgtttt acttgtacaa
      661 ctaataataa cttagttatt tagagaattt cacttgttga ctagaaaaat aatttaatat
      721 tgaaacaatt gcaccataaa ttgtataaat ataaaattaa ttctaattgt tttttttatt
      781 gcagtttctt gccgcgggaa ttcgattact agtatgccac ccaagcgcaa aacgcttaac
      841 gctgcggctg aggctaacgc tcatgccgac ggacacgccg acggaaacgc cgacggacac
      901 gtggccaata cggccgcgtc ctcgaataat gcgaggttcg ctgatctcac taacatcgat
      961 actccgggtc tgggacccac aactacgacc ctgctcgtgg aaccagcacg ctcaaagcgt
     1021 caacgagtgt cccgcgcatg cgaccagtgc cgtgcagccc gagagaaatg cgacggaata
     1081 cagcctgcgt gtttcccgtg cgtttcccag ggaaggtcct gcacttatca ggcttcgccg
     1141 aaaaagaggg gagttcaaac cggttatatt cgtacgctgg agctcgccct cgcctggatg
     1201 tttgaaaatg tcgcgcgttc cgaagatgcc ttgcataacc tcctcgtccg tgacgccgga
     1261 caaggatcag ctctgctcgt tggtaaagat tcgccggctg ccgagcgact ccatgcccgt
     1321 tgggctacta gccgtgtcaa taagagcatt acccgcctcc tccgtcagtt ggagctccct
     1381 cctaccgcca cggctacggc ctcgataatg ccgcacgtga tggagcagcc tctcagtacc
     1441 agcattaacc ccgtcaacga ccgcttcaac ggtattccca accccactcc gtataactcc
     1501 gatgcagctc tcgatgctat cactcagacc aacgattatg gaagcgtaaa tacacatggt
     1561 atcctctcta cttacccgcc accggctacg caccttaatg aagcttccgt cgctctcgct
     1621 cccggtggcg cccccccccg accgcctcct ccgtatgttg acagcacgac caatcacccg
     1681 ccgtaccact cgaatctggt tccaatggcg aactttggtt actcgaccgt tgattacgat
     1741 gccatggttg acgatttggc tagcattgaa tacacggacg ctgtggatgt cgacccacag
     1801 tttatgacca atctgggatt cgttcctgga tgtaacttct ccgacattaa tacatacgaa
     1861 cagtgatgac gtctatcgat accgtcgact aaagccaacc cagctttctt gtacaaagtg
     1921 ggggatccag acatgataag atacattgat gagtttggac aaaccacaac tagagtgcag
     1981 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     2041 agctgcaata aacaagttaa caataacaat tgcattcatt ttatgtttca ggttcagggg
     2101 gaggtgtggg aggttttttt caactttatt atacatagtt gctgcagaat cactagtgaa
     2161 ttcgcggccg cctgcaggtc gaccatatgg gagagctccc aacgcgttgg atgcatagct
     2221 tgagtattct atagtgtcac ctaaatagct tggcgtaatc atggtcatag ctgtttcctg
     2281 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2341 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2401 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     2461 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     2521 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     2581 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     2641 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     2701 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     2761 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     2821 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     2881 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     2941 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3001 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3061 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3121 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3181 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3241 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3301 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3361 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3421 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3481 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     3541 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     3601 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     3661 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     3721 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     3781 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     3841 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     3901 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     3961 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4021 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4081 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4141 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4201 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4261 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4321 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4381 gggttccgcg cacatttccc cgaaaagtgc cacctgatgc ggtgtgaaat accgcacaga
     4441 tgcgtaagga gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg
     4501 cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc
     4561 cttataaatc aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga
     4621 gtccactatt aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg
     4681 atggcccact acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag
     4741 cactaaatcg gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga
     4801 acgtggcgag aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg
     4861 tagcggtcac gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg
     4921 cgtccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc
     4981 gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc
     5041 agggttttcc cagtcac
//
