LOCUS       pWHERE.        9944 bp DNA     circular SYN 06-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    pWHERE.
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9944)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9944)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9944
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    complement(1..2032)
                     /label=H19 ins
                     /note="H19 ins"
                     /note="mH19 insulators on either side of the lacZ
                     transcription unit. Both insulators are expected to protect
                     the integrated transcriptional LacZ unit from negative as 
                     well as positive influences from neighboring sequences. 
                     Insulator elements can be functionally identified by their 
                     ability to shield promoters from regulators in a 
                     position-dependent manner or by their ability to protect 
                     adjacent transgenes from position effects. The fragment of 
                     the differential methylated region (DMD) located between 
                     the mouse Igf2 and H19 acts as a powerful insulator. The 
                     enhancer blocking activity of the DMD fragment is dependent
                     upon four responsive elements to the vertebrate 
                     enhancer-blocking protein CTCF. Two mouse DMD fragments 
                     have been introduced in opposite orientation in the pWHERE 
                     plasmid to insulate your promoter of interest cloned 
                     upstream of the new CpG-free LacZ gene from the 5' and 3' 
                     adjacent regions at the integrated site in transgenic 
                     mice."
     CDS             2302..2322
                     /label=SV40 NLS
                     /note="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
     polyA_signal    5170..5742
                     /label=EF-1-alpha poly(A) signal
                     /note="human EF-1-alpha polyadenylation signal"
     misc_feature    complement(5750..7781)
                     /label=H19 ins
                     /note="H19 ins"
                     /note="mH19 insulators on either side of the lacZ
                     transcription unit. Both insulators are expected to protect
                     the integrated transcriptional LacZ unit from negative as 
                     well as positive influences from neighboring sequences. 
                     Insulator elements can be functionally identified by their 
                     ability to shield promoters from regulators in a 
                     position-dependent manner or by their ability to protect 
                     adjacent transgenes from position effects. The fragment of 
                     the differential methylated region (DMD) located between 
                     the mouse Igf2 and H19 acts as a powerful insulator. The 
                     enhancer blocking activity of the DMD fragment is dependent
                     upon four responsive elements to the vertebrate 
                     enhancer-blocking protein CTCF. Two mouse DMD fragments 
                     have been introduced in opposite orientation in the pWHERE 
                     plasmid to insulate your promoter of interest cloned 
                     upstream of the new CpG-free LacZ gene from the 5' and 3' 
                     adjacent regions at the integrated site in transgenic 
                     mice."
     rep_origin      complement(7877..8465)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8639..9496)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(9497..9601)
                     /label=AmpR promoter
ORIGIN
        1 cagctttgtc acagcggacc ccaacctatg ccgcgtctgc cgagcaatat gtagtattgt
       61 actgccacca cgcggcatcg tctgtccatt tagctatagc caaatctgca cagcgtggag
      121 agtgaaccga tcgatcgtga agggcgcaag actgaaggag ctacccaaga aatgtgtgtt
      181 gtaccacccc atgaccctta tgaatcattg aatgctatgc ctgagtgacc catgagtttg
      241 ccataggtga accgcaattt tggtcacctc tatgataaag tatgtgggac tgcactggtc
      301 gctgctcggc aacttcggtc ttaccagcca ctgacgatct cgggctgtgt agggaatgag
      361 tcaagttctc tggttcagtg tgtaagggaa ccattccaga ggtgcacaca tcttaccacc
      421 cctatgaatc cctatttggg tgaccctggg atattgctgg gaatgaatcg ctccccccaa
      481 gttggcagca tttgggccac gatatatagg agtatgctgc caccgcgcgg tagcatccgt
      541 tcccttgttg cacataacag cttctatgcc ttcctatagt gagccacact ggctggtttt
      601 ggggttcagt gaccaaaggg accccctcca gaaacacaag tgttccaacc tcttataaac
      661 catatcgacc actgaggcat agcggcttcg gacattgctg tgggcgaacc cgaactttgg
      721 cccttggaca ttgtcatggg caaaccttaa ctttggctgt ctctggaaca taatgtttaa
      781 gatgacagtc accagcgcag caatttggtc tttccactca caacggcttt tgtgctttct
      841 ggcatcgaac cacatgcact ggtttatggg gtctgagacc aaaggagacc attccggaag
      901 ggcataggtg tcctgccttc tgcttttaac aaggctctcc gggacagtgc aaaacaggtg
      961 aaccccaact ttgccataag tacgattatc tgccacaaac cagccagggg tcctaccacc
     1021 tcttcaattg attttgggct gacacccaag gcttgatgta ggattcctca gctgccaagc
     1081 tggcagctga cccccattga gagagaatgc agtttcagaa ttgtttgcag ccctgagccg
     1141 gagatcatta gcatctgaac gccccaatta gaatacgaaa gcgcaaatca ccagactttc
     1201 ttgttggcgg ttccttaagt gattctttgg gtagggaggt caggtgtcct catagatgtg
     1261 cagaatttga ggaccatgct tagtggggtc tgcattatgg tactgcaata cattccatga
     1321 tcaccacaca tagtagctat acttcaattt tcacacaata gcgctgatgg ccccagaacc
     1381 ctataagtca gatacctgag atagctcttg agaacgtttt atcaaggact agcatgaacc
     1441 cctggcctca tgaagcccat gactatggga tcatagatgg tgatagggga gaaaactcaa
     1501 tcagttgcaa tccgttttag gactgcgatg tacgagactt cactgccgcc gtgcggcaac
     1561 cctggtcttt acacacaaag gattctttgc agagagtaag ccgaccttgt tgatttggga
     1621 gtccgagtcc acgaggtacc agcctagaaa atgcatgtgt cctgcccccc tagtggcatt
     1681 tgtgaccccc ctgaggtact gaacttgggt gacccacagc attgccattt gtgaattcca
     1741 ataccagggg tgggggggct ctttaggttt ggcgcaatcg attttgctgc caccacgcgg
     1801 caactcccgc gtataaaccc cacaactgat tcagcagacg tccaagaata gggcatggtc
     1861 tccttgcaga attcttatgc ctcctggatg ctcgtgtgaa tgtagcatgt tcctttgagt
     1921 cctgggtgta agtgccctgc acgctcatcc cccggacatg aaaatagaat ctctattttc
     1981 tcaccaacct ttttctttcc ttgtggtgat tcgggaactg taggcaatgg ctcctgcagg
     2041 tcctaggtgg atcctctcga gtcccgggcc atggaccctg ttgtgctgca aaggagagac
     2101 tgggagaacc ctggagtgac ccagctcaac agactggctg cccaccctcc ctttgcctct
     2161 tggaggaact ctgaggaagc caggacagac aggcccagcc agcagctcag gtctctcaat
     2221 ggagagtgga ggtttgcctg gttccctgcc cctgaagctg tgcctgagtc ttggctggag
     2281 tgtgacctcc cagaggcagt tcccaagaag aagaggaaag ttgaggctga cactgttgtg
     2341 gtgccaagca actggcagat gcatggctat gatgccccca tctacaccaa tgtcacctac
     2401 cccatcactg tgaacccccc ttttgtgccc actgagaacc ccactggctg ctacagcctg
     2461 accttcaatg ttgatgagag ctggctgcaa gaaggccaga ccaggatcat ctttgatgga
     2521 gtcaactctg ccttccacct ctggtgcaat ggcaggtggg ttggctatgg ccaagacagc
     2581 aggctgccct ctgagtttga cctctctgcc ttcctcagag ctggagagaa caggctggct
     2641 gtcatggtgc tcaggtggtc tgatggcagc tacctggaag accaagacat gtggaggatg
     2701 tctggcatct tcagggatgt gagcctgctg cacaagccca ccacccagat ttctgacttc
     2761 catgttgcca ccaggttcaa tgatgacttc agcagagctg tgctggaggc tgaggtgcag
     2821 atgtgtggag aactcagaga ctacctgaga gtcacagtga gcctctggca aggtgagacc
     2881 caggtggcct ctggcacagc cccctttgga ggagagatca ttgatgagag aggaggctat
     2941 gctgacagag tcaccctgag gctcaatgtg gagaacccca agctgtggtc tgctgagatc
     3001 cccaacctct acagggctgt tgtggagctg cacactgctg atggcaccct gattgaagct
     3061 gaagcctgtg atgttggatt cagagaagtc aggattgaga atggcctgct gctgctcaat
     3121 ggcaagcctc tgctcatcag gggagtcaac aggcatgagc accaccctct gcatggacaa
     3181 gtgatggatg aacagacaat ggtgcaagat atcctgctaa tgaagcagaa caacttcaat
     3241 gctgtcaggt gctctcacta ccccaaccac cctctctggt acaccctgtg tgacaggtat
     3301 ggcctgtatg ttgttgatga agccaacatt gagacacatg gcatggtgcc catgaacagg
     3361 ctcacagatg accccaggtg gctgcctgcc atgtctgaga gagtgaccag gatggtgcag
     3421 agagacagga accacccctc tgtgatcatc tggtctctgg gcaatgagtc tggacatgga
     3481 gccaaccatg atgctctcta caggtggatc aagtctgttg accccagcag acctgtgcag
     3541 tatgaaggag gtggagcaga caccacagcc acagacatca tctgccccat gtatgccagg
     3601 gttgatgagg accagccctt ccctgctgtg cccaagtgga gcatcaagaa gtggctctct
     3661 ctgcctggag agaccagacc tctgatcctg tgtgaatatg cacatgcaat gggcaactct
     3721 ctgggaggct ttgccaagta ctggcaagcc ttcagacagt accccaggct gcaaggagga
     3781 tttgtgtggg actgggtgga ccaatctctc atcaagtatg atgagaatgg caacccctgg
     3841 tctgcctatg gaggagactt tggtgacacc cccaatgaca ggcagttctg catgaatggc
     3901 ctggtctttg cagacaggac ccctcaccct gccctcacag aggccaagca ccagcaacag
     3961 ttcttccagt tcaggctgtc tggacagacc attgaggtga catctgagta cctcttcagg
     4021 cactctgaca atgagctcct gcactggatg gtggccctgg atggcaagcc tctggcttct
     4081 ggtgaggtgc ctctggatgt ggcccctcaa ggaaagcagc tgattgaact gcctgagctg
     4141 cctcagccag agtctgctgg acaactgtgg ctaacagtga gggtggttca gcccaatgca
     4201 acagcttggt ctgaggcagg ccacatctct gcatggcagc agtggaggct ggctgagaac
     4261 ctctctgtga ccctgcctgc tgcctctcat gccatccctc acctgacaac atctgaaatg
     4321 gacttctgca ttgagctggg caacaagaga tggcagttca acaggcagtc tggcttcctg
     4381 tctcagatgt ggattggaga caagaagcag ctcctcaccc ctctcaggga ccaattcacc
     4441 agggctcctc tggacaatga cattggagtg tctgaggcca ccaggattga cccaaatgct
     4501 tgggtggaga ggtggaaggc tgctggacac taccaggctg aggctgccct gctccagtgc
     4561 acagcagaca ccctggctga tgctgttctg atcaccacag cccatgcttg gcagcaccaa
     4621 ggcaagaccc tgttcatcag cagaaagacc tacaggattg atggctctgg acagatggca
     4681 atcacagtgg atgtggaggt tgcctctgac acacctcacc ctgcaaggat tggcctgaac
     4741 tgtcaactgg cacaggtggc tgagagggtg aactggctgg gcttaggccc tcaggagaac
     4801 taccctgaca ggctgacagc tgcctgcttt gacaggtggg acctgcctct gtctgacatg
     4861 tacacccctt atgtgttccc ttctgagaat ggcctgaggt gtggcaccag ggagctgaac
     4921 tatggtcctc accagtggag gggagacttc cagttcaaca tctccaggta ctctcagcaa
     4981 cagctcatgg aaacctctca caggcacctg ctccatgcag aggagggaac ctggctgaac
     5041 attgatggct tccacatggg cattggagga gatgactctt ggtctccttc tgtgtctgct
     5101 gagttccagt tatctgctgg caggtaccac tatcagctgg tgtggtgcca gaagtaaacc
     5161 tgagctagca ttatccctaa tacctgccac cccactctta atcagtggtg gaagaacggt
     5221 ctcagaactg tttgtttcaa ttggccattt aagtttagta gtaaaagact ggttaatgat
     5281 aacaatgcat cgtaaaacct tcagaaggaa aggagaatgt tttgtggacc actttggttt
     5341 tcttttttgc gtgtggcagt tttaagttat tagtttttaa aatcagtact ttttaatgga
     5401 aacaacttga ccaaaaattt gtcacagaat tttgagaccc attaaaaaag ttaaatgaga
     5461 aacctgtgtg ttcctttggt caacaccgag acatttaggt gaaagacatc taattctggt
     5521 tttacgaatc tggaaacttc ttgaaaatgt aattcttgag ttaacacttc tgggtggaga
     5581 atagggttgt tttcccccca cataattgga aggggaagga atatcattta aagctatggg
     5641 agggtttctt tgattacaac actggagaga aatgcagcat gttgctgatt gcctgtcact
     5701 aaaacaggcc aaaaactgag tccttgggtt gcatagaaag ctgcctgcag agctttgtca
     5761 cagcggaccc caacctatgc cgcgtctgcc gagcaatatg tagtattgta ctgccaccac
     5821 gcggcatcgt ctgtccattt agctatagcc aaatctgcac agcgtggaga gtgaaccgat
     5881 cgatcgtgaa gggcgcaaga ctgaaggagc tacccaagaa atgtgtgttg taccacccca
     5941 tgacccttat gaatcattga atgctatgcc tgagtgaccc atgagtttgc cataggtgaa
     6001 ccgcaatttt ggtcacctct atgataaagt atgtgggact gcactggtcg ctgctcggca
     6061 acttcggtct taccagccac tgacgatctc gggctgtgta gggaatgagt caagttctct
     6121 ggttcagtgt gtaagggaac cattccagag gtgcacacat cttaccaccc ctatgaatcc
     6181 ctatttgggt gaccctggga tattgctggg aatgaatcgc tccccccaag ttggcagcat
     6241 ttgggccacg atatatagga gtatgctgcc accgcgcggt agcatccgtt cccttgttgc
     6301 acataacagc ttctatgcct tcctatagtg agccacactg gctggttttg gggttcagtg
     6361 accaaaggga ccccctccag aaacacaagt gttccaacct cttataaacc atatcgacca
     6421 ctgaggcata gcggcttcgg acattgctgt gggcgaaccc gaactttggc ccttggacat
     6481 tgtcatgggc aaaccttaac tttggctgtc tctggaacat aatgtttaag atgacagtca
     6541 ccagcgcagc aatttggtct ttccactcac aacggctttt gtgctttctg gcatcgaacc
     6601 acatgcactg gtttatgggg tctgagacca aaggagacca ttccggaagg gcataggtgt
     6661 cctgccttct gcttttaaca aggctctccg ggacagtgca aaacaggtga accccaactt
     6721 tgccataagt acgattatct gccacaaacc agccaggggt cctaccacct cttcaattga
     6781 ttttgggctg acacccaagg cttgatgtag gattcctcag ctgccaagct ggcagctgac
     6841 ccccattgag agagaatgca gtttcagaat tgtttgcagc cctgagccgg agatcattag
     6901 catctgaacg ccccaattag aatacgaaag cgcaaatcac cagactttct tgttggcggt
     6961 tccttaagtg attctttggg tagggaggtc aggtgtcctc atagatgtgc agaatttgag
     7021 gaccatgctt agtggggtct gcattatggt actgcaatac attccatgat caccacacat
     7081 agtagctata cttcaatttt cacacaatag cgctgatggc cccagaaccc tataagtcag
     7141 atacctgaga tagctcttga gaacgtttta tcaaggacta gcatgaaccc ctggcctcat
     7201 gaagcccatg actatgggat catagatggt gataggggag aaaactcaat cagttgcaat
     7261 ccgttttagg actgcgatgt acgagacttc actgccgccg tgcggcaacc ctggtcttta
     7321 cacacaaagg attctttgca gagagtaagc cgaccttgtt gatttgggag tccgagtcca
     7381 cgaggtacca gcctagaaaa tgcatgtgtc ctgcccccct agtggcattt gtgacccccc
     7441 tgaggtactg aacttgggtg acccacagca ttgccatttg tgaattccaa taccaggggt
     7501 gggggggctc tttaggtttg gcgcaatcga ttttgctgcc accacgcggc aactcccgcg
     7561 tataaacccc acaactgatt cagcagacgt ccaagaatag ggcatggtct ccttgcagaa
     7621 ttcttatgcc tcctggatgc tcgtgtgaat gtagcatgtt cctttgagtc ctgggtgtaa
     7681 gtgccctgca cgctcatccc ccggacatga aaatagaatc tctattttct caccaacctt
     7741 tttctttcct tgtggtgatt cgggaactgt aggcaatggc tttaattaaa aacccgcttc
     7801 ggcgggtttt tttatgcatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     7861 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     7921 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     7981 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     8041 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     8101 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     8161 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     8221 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     8281 tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc
     8341 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     8401 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     8461 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     8521 gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt
     8581 aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc
     8641 aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg
     8701 cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg
     8761 ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc
     8821 cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta
     8881 ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg
     8941 ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct
     9001 ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta
     9061 gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg
     9121 ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga
     9181 ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt
     9241 gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca
     9301 ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt
     9361 cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt
     9421 ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga
     9481 aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat cagggttatt
     9541 gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc
     9601 gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa
     9661 cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt gatgacggtg
     9721 aaaacctctg acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     9781 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     9841 actatgcggc atcagagcag attgtactga gagtgcacca tatggatctc gataacaaaa
     9901 aaccccgccc cggcggggtt ttttgttagc ggccgcttaa ttaa
//
