LOCUS       sequence-459307-        7121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of FKBP-SPO11.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7121)
  TITLE     Keeney Lab SPO11 plasmids
REFERENCE   2  (bases 1 to 7121)
  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"
COMMENT     Sequence Label: pZZMP152
FEATURES             Location/Qualifiers
     source          1..7121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      901..910
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             910..933
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             934..1254
                     /codon_start=1
                     /product="human FK506-binding protein FKBP12"
                     /label=FKBP
                     /translation="GVQVETISPGDGRTFPKRGQTCVVHYTGMLEDGKKFDSSRDRNKP
                     FKFMLGKQEVIRGWEEGVAQMSVGQRAKLTISPDYAYGATGHPGIIPPHATLVFDVELL
                     KLE"
     CDS             1297..1317
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(2543..2560)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2549..2773
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2819..3247
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2906..2925)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3116..3137
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3256..3276)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3261..3590
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3441..3576
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3503..3522
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3639..4664
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     CDS             3639..4664
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     E"
     polyA_signal    4794..4915
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4831..4850)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4885..4904
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4964..4980)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4964..4980)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4977..4999)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4988..5004
                     /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(5012..5018,5019..5036,5037..5042))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5057..5078
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5195..5212)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5366..5954)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5446..5465)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6125..6916,6917..6985))
                     /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     6748..6767
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6986..7090)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gccaccatgg attacaagga tgacgacgat aagggcgttc aagtggaaac tataagccca
      961 ggagatggtc gcacattccc gaaacgcggt caaacttgcg tggtgcatta tacagggatg
     1021 cttgaggacg gtaagaaatt tgacagttct agggatcgga acaaaccgtt taagttcatg
     1081 ctgggtaagc aagaggttat acggggatgg gaagagggcg tcgcgcagat gagtgtcgga
     1141 cagcgggcca agttgacaat ctcacctgac tatgcatacg gtgctacggg gcatcctggg
     1201 atcattcccc ctcatgccac tctggttttt gacgttgaac ttctcaaatt ggagggttcc
     1261 ggtagcggag ggggtagcgg tggctctggg ggatctgaga acctctactt ccagggagct
     1321 tttgccccca tgggacctga ggccagcttc ttcgatgccc tggatagaca tagagcctct
     1381 ctgctggcca tggtgaaaag aggcgctggc gagacccctg ctggcgccac aagagtcgcc
     1441 tccagctccg aggtgctgac agccatcgaa aacatcatcc aggacatcat caagtccctg
     1501 gctagaaacg aggtgcctgc cttcaccatc gacaacagat ctagctggga gaacatcatg
     1561 ttcgacgata gtgtgggcct gcggatgatc ccccagtgca ccaccagaaa gatcagaagc
     1621 gacagcccta agagcgtgaa gaaattcgcc ctgattctga aggtgctgag catgatctac
     1681 aagctgatcc agagcgacac ctacgccacc aagcgggaca tctactacac cgactcccag
     1741 ctgttcggca accaggccgc cgtggattct gccatcgatg acatcagctg catgctgaaa
     1801 gtgcccagac ggagcctgca cgtgttgtct acaagcaagg gcctgatcgc cggcaatctg
     1861 cggtacatgg aagaggacgg cacccgggtg cagtgtacct gcagcgccac cgccacagcc
     1921 gtgcctacga acatccaggg catgcagcac ctgatcaccg acgccaagtt cctgctgatc
     1981 gtggaaaaag atgctacctt ccagcggctg ctggacgaca acttctgcag cagaatgagc
     2041 ccttgtatca tggtcacagg caagggcgtg ccagacctga acacccggct gctggtgaag
     2101 aagctgtggg acacatttca catcccggtt ttcaccctgg tggacgccga tccttacgga
     2161 atcgagatca tgtgcatcta taagtacggc agcatgagca tgagctttga ggcccacaat
     2221 ctgacaattc caacaattcg gtggctggga ctgctgccca gcgatatcca aagacttaat
     2281 atccccaagg acagcctgat ccctctgacc aagcacgacc agatgaaact ggacagcatc
     2341 ctgaagagac cttatatcac ctaccagcct ctctggaaga aggaactgga aatgatggcc
     2401 gactctaaga tgaaggccga gatccaagct ctgacactac tgagctctga ttacctgtct
     2461 agagtgtacc tgcctaacaa gctgaggttc ggcggatgga tctaactcga gtctagaggg
     2521 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     2581 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     2641 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     2701 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     2761 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     2821 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     2881 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     2941 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     3001 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     3061 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     3121 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     3181 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     3241 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     3301 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     3361 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     3421 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     3481 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     3541 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     3601 gagcttgtat atccattttc ggatctgatc agcacgtgat gaaaaagcct gaactcaccg
     3661 cgacgtctgt cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc
     3721 tctcggaggg cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc
     3781 tgcgggtaaa tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg
     3841 catcggccgc gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga
     3901 cctattgcat ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac
     3961 tgcccgctgt tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta
     4021 gccagacgag cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc
     4081 gtgatttcat atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg
     4141 acaccgtcag tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact
     4201 gccccgaagt ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca
     4261 atggccgcat aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg
     4321 aggtcgccaa catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct
     4381 acttcgagcg gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc
     4441 gcattggtct tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt
     4501 gggcgcaggg tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac
     4561 aaatcgcccg cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata
     4621 gtggaaaccg acgccccagc actcgtccga gggcaaagga atagcacgtg ctacgagatt
     4681 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4741 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4801 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4861 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4921 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     4981 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5041 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5101 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5161 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5221 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5281 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5341 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5401 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5461 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5521 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5581 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5641 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5701 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5761 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     5821 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     5881 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     5941 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6001 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6061 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6121 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6181 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6241 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6301 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6361 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6421 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6481 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6541 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6601 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6661 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6721 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6781 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     6841 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     6901 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     6961 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7021 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7081 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt c
//
