LOCUS       40924_33542        7104 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pNT1.1 for cKO, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7104)
  AUTHORS   Tokuhiro K, Ikawa M, Benham AM, Okabe M.
  TITLE     Protein disulfide isomerase homolog PDILT is required for quality 
            control of sperm membrane protein ADAM3 and male fertility 
            [corrected]
  JOURNAL   Proc. Natl. Acad. Sci. U.S.A. 109 (10), 3850-3855 (2012)
  PUBMED    22357757
REFERENCE   2  (bases 1 to 7104)
  AUTHORS   Ikawa M.
  TITLE     Direct Submission
  JOURNAL   Submitted (26-OCT-2011) Rimd, Osaka University, 3-1 Yamada-Oka, 
            Suita, Osaka 5650871, Japan
REFERENCE   3  (bases 1 to 7104)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7104)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc. Natl.
            Acad. Sci. U.S.A."; date: "2012"; volume: "109"; issue: "10"; pages:
            "3850-3855"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (26-OCT-2011) Rimd, Osaka University, 3-1 Yamada-Oka, Suita, Osaka 
            5650871, Japan"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7104
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    7..61
                     /note="multi-cloning site 1 (PmeI, NotI, SalI, NsiI, AsiSI,
                     XhoI, SfiI)"
     misc_feature    91..124
                     /label=LoxP site 1
                     /note="LoxP site 1"
     protein_bind    91..124
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     misc_feature    137..142
                     /label=BamHI
                     /note="BamHI"
     misc_feature    146..193
                     /label=FRT site 1
                     /note="FRT site 1"
     protein_bind    complement(146..193)
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     polyA_signal    complement(203..251)
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     CDS             complement(261..1061)
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     promoter        complement(1075..1574)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     misc_feature    1601..1606
                     /label=HindIII
                     /note="HindIII"
     protein_bind    complement(1610..1657)
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    1661..1686
                     /label=multi-cloning site 2 (EcoRI, NheI, AflII, AscI)
                     /note="multi-cloning site 2 (EcoRI, NheI, AflII, AscI)"
     protein_bind    complement(1687..1720)
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     misc_feature    1721..1750
                     /label=multi-cloning site 3 (PacI, KpnI, MfeI, AvrII)
                     /note="multi-cloning site 3 (PacI, KpnI, MfeI, AvrII)"
     promoter        1765..2264
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     5'UTR           2306..2345
                     /label=HSV TK 5'-UTR
                     /note="5' untranslated region from the herpes simplex virus
                     thymidine kinase gene"
     CDS             2363..3490
                     /label=HSV TK
                     /note="herpes simplex virus thymidine kinase"
     polyA_signal    3535..3583
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     polyA_signal    4100..4554
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation
                     signal"
     misc_feature    4558..4563
                     /label=ClaI
                     /note="ClaI"
     promoter        4927..5031
                     /label=AmpR promoter
     CDS             5032..5889
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      6063..6651
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    6939..6960
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        6975..7005
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7013..7029
                     /label=lac operator
                     /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)."
     primer_bind     7037..7053
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        7074..7092
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctcgttt aaacgcggcc gcgtcgacat gcatgcgatc gcctcgaggg ccgcaatggc
       61 cagtactagt gaacctcttc gagggaccta ataacttcgt atagcataca ttatacgaag
      121 ttatattaag ggttccggat ccgatgaagt tcctatactt tctagagaat aggaacttcg
      181 gaataggaac ttcatcagat ccgaacaaac gacccaacac ccgtgcgttt tattctgtct
      241 ttttattgcc gatcccctca gaagaactcg tcaagaaggc gatagaaggc gatgcgctgc
      301 gaatcgggag cggcgatacc gtaaagcacg aggaagcggt cagcccattc gccgccaagc
      361 tcttcagcaa tatcacgggt agccaacgct atgtcctgat agcggtccgc cacacccagc
      421 cggccacagt cgatgaatcc agaaaagcgg ccattttcca ccatgatatt cggcaagcag
      481 gcatcgccat gggtcacgac gagatcctcg ccgtcgggca tgcgcgcctt gagcctggcg
      541 aacagttcgg ctggcgcgag cccctgatgc tcttcgtcca gatcatcctg atcgacaaga
      601 ccggcttcca tccgagtacg tgctcgctcg atgcgatgtt tcgcttggtg gtcgaatggg
      661 caggtagccg gatcaagcgt atgcagccgc cgcattgcat cagccatgat ggatactttc
      721 tcggcaggag caaggtgaga tgacaggaga tcctgccccg gcacttcgcc caatagcagc
      781 cagtcccttc ccgcttcagt gacaacgtcg agcacagctg cgcaaggaac gcccgtcgtg
      841 gccagccacg atagccgcgc tgcctcgtcc tgcagttcat tcagggcacc ggacaggtcg
      901 gtcttgacaa aaagaaccgg gcgcccctgc gctgacagcc ggaacacggc ggcatcagag
      961 cagccgattg tctgttgtgc ccagtcatag ccgaatagcc tctccaccca agcggccgga
     1021 gaacctgcgt gcaatccatc ttgttcaatg gccgatccca tattggctgc aggtcgaaag
     1081 gcccggagat gaggaagagg agaacagcgc ggcagacgtg cgcttttgaa gcgtgcagaa
     1141 tgccgggcct ccggaggacc ttcgggcgcc cgccccgccc ctgagcccgc ccctgagccc
     1201 gcccccggac ccaccccttc ccagcctctg agcccagaaa gcgaaggagc aaagctgcta
     1261 ttggccgctg ccccaaaggc ctacccgctt ccattgctca gcggtgctgt ccatctgcac
     1321 gagactagtg agacgtgcta cttccatttg tcacgtcctg cacgacgcga gctgcggggc
     1381 gggggggaac ttcctgacta ggggaggagt agaaggtggc gcgaaggggc caccaaagaa
     1441 cggagccggt tggcgcctac cggtggatgt ggaatgtgtg cgaggccaga ggccacttgt
     1501 gtagcgccaa gtgcccagcg gggctgctaa agcgcatgct ccagactgcc ttgggaaaag
     1561 cgcctcccct acccggtaga attgacctgc aggggcccct aagcttgtcg aagttcctat
     1621 actttctaga gaataggaac ttcggaatag gaacttcgac gaattcgcta gccttaaggg
     1681 cgcgccataa cttcgtatag catacattat acgaagttat ttaattaagg taccaacaat
     1741 tgttcctagg ctagaaattc taccgggtag gggaggcgct tttcccaagg cagtctggag
     1801 catgcgcttt agcagccccg ctgggcactt ggcgctacac aagtggcctc tggcctcgca
     1861 cacattccac atccaccggt aggcgccaac cggctccgtt ctttggtggc cccttcgcgc
     1921 caccttctac tcctccccta gtcaggaagt tcccccccgc cccgcagctc gcgtcgtgca
     1981 ggacgtgaca aatggaagta gcacgtctca ctagtctcgt gcagatggac agcaccgctg
     2041 agcaatggaa gcgggtaggc ctttggggca gcggccaata gcagctttgc tccttcgctt
     2101 tctgggctca gaggctggga aggggtgggt ccgggggcgg gctcaggggc gggctcaggg
     2161 gcggggcggg cgcccgaagg tcctccggag gcccggcatt ctgcacgctt caaaagcgca
     2221 cgtctgccgc gctgttctcc tcttcctcat ctccgggcct ttcgacctgc agcgacccgc
     2281 ttaacagcgt caacagcgtg ccgcagatct tggtggcgtg aaactcccgc acctcttcgg
     2341 caagcgcctt gtagaagcgc gtatggcttc gtacccctgc catcaacacg cgtctgcgtt
     2401 cgaccaggct gcgcgttctc gcggccatag caaccgacgt acggcgttgc gccctcgccg
     2461 gcagcaagaa gccacggaag tccgcctgga gcagaaaatg cccacgctac tgcgggttta
     2521 tatagacggt cctcacggga tggggaaaac caccaccacg caactgctgg tggccctggg
     2581 ttcgcgcgac gatatcgtct acgtacccga gccgatgact tactggcagg tgctgggggc
     2641 ttccgagaca atcgcgaaca tctacaccac acaacaccgc ctcgaccagg gtgagatatc
     2701 ggccggggac gcggcggtgg taatgacaag cgcccagata acaatgggca tgccttatgc
     2761 cgtgaccgac gccgttctgg ctcctcatat cgggggggag gctgggagct cacatgcccc
     2821 gcccccggcc ctcaccctca tcttcgaccg ccatcccatc gccgccctcc tgtgctaccc
     2881 ggccgcgcga taccttatgg gcagcatgac cccccaggcc gtgctggcgt tcgtggccct
     2941 catcccgccg accttgcccg gcacaaacat cgtgttgggg gcccttccgg aggacagaca
     3001 catcgaccgc ctggccaaac gccagcgccc cggcgagcgg cttgacctgg ctatgctggc
     3061 cgcgattcgc cgcgtttacg ggctgcttgc caatacggtg cggtatctgc agggcggcgg
     3121 gtcgtggcgg gaggattggg gacagctttc ggggacggcc gtgccgcccc agggtgccga
     3181 gccccagagc aacgcgggcc cacgacccca tatcggggac acgttattta ccctgtttcg
     3241 ggcccccgag ttgctggccc ccaacggcga cctgtacaac gtgtttgcct gggccttgga
     3301 cgtcttggcc aaacgcctcc gtcccatgca cgtctttatc ctggattacg accaatcgcc
     3361 cgccggctgc cgggacgccc tgctgcaact tacctccggg atggtccaga cccacgtcac
     3421 cacccccggc tccataccga cgatctgcga cctggcgcgc acgtttgccc gggagatggg
     3481 ggaggctaac tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa
     3541 taaaaagaca gaataaaacg cacgggtgtt gggtcgtttg ttcataaacg cggggttcgg
     3601 tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc
     3661 gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc
     3721 aacgtcgggg cggcaagccc tgccatagcc acgggccccg tgggttaggg acggggtccc
     3781 ccatggggaa tggtttatgg ttcgtggggg ttattatttt gggcgttgcg tggggtcagg
     3841 tccacgactg gactgagcag acagacccat ggtttttgga tggcctgggc atggaccgca
     3901 tgtactggcg cgacacgaac accgggcgtc tgtggctgcc aaacaccccc gacccccaaa
     3961 aaccaccgcg cggatttctg gcgccgccgg acgaactaaa cctgactacg gcatctctgc
     4021 cccttcttcg ctggtacgag gagcgctttt gttttgtatt ggtcaccacg gccgagtttc
     4081 cgcgggaccc cggccaggac ctgcagaaat tgatgatcta ttaaacaata aagatgtcca
     4141 ctaaaatgga agtttttcct gtcatacttt gttaagaagg gtgagaacag agtacctaca
     4201 ttttgaatgg aaggattgga gctacggggg tgggggtggg gtgggattag ataaatgcct
     4261 gctctttact gaaggctctt tactattgct ttatgataat gtttcatagt tggatatcat
     4321 aatttaaaca agcaaaacca aattaagggc cagctcattc ctcccactca tgatctatag
     4381 atctatagat ctctcgtggg atcattgttt ttctcttgat tcccactttg tggttctaag
     4441 tactgtggtt tccaaatgtg tcagtttcat agcctgaaga acgagatcag cagcctctgt
     4501 tccacataca cttcattctc agtattgttt tgccaagttc taattccatc agaagctatc
     4561 gatgtcgagg gggggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4621 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4681 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4741 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4801 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4861 atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac ttttcgggga
     4921 aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     4981 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     5041 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     5101 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     5161 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     5221 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     5281 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     5341 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     5401 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     5461 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     5521 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     5581 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     5641 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     5701 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     5761 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     5821 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     5881 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     5941 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     6001 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     6061 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     6121 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     6181 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     6241 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     6301 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     6361 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     6421 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     6481 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     6541 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     6601 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     6661 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     6721 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     6781 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca
     6841 atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg
     6901 tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat
     6961 taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc
     7021 ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagcg cgcaattaac
     7081 cctcactaaa gggaacaaaa gctg
//
