LOCUS       40924_515        6228 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Mammalian vector Cre-excised pCOIN DV, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6228)
  AUTHORS   De Schamphelaire W, Olbrechts A, Meert J, Verhelst K, Roggeman 
            Fonseca M, Vanhoucke M, Beyaert R.
  TITLE     BCCM/LMBP Plasmid collection
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 6228)
  AUTHORS   De Schamphelaire W.
  TITLE     Direct Submission
  JOURNAL   Submitted (02-FEB-2017) BCCM/LMBP, Universiteit Gent, 
            Technologiepark 927, 9052, BELGIUM
REFERENCE   3  (bases 1 to 6228)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 6228)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (02-FEB-2017) BCCM/LMBP, Universiteit Gent, Technologiepark 927, 
            9052, BELGIUM"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6228
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    35..277
                     /label=chicken beta-globin 5'HS4 insulator core sequence
                     /note="chicken beta-globin 5'HS4 insulator core sequence"
     misc_feature    278..520
                     /label=chicken beta-globin 5'HS4 insulator core sequence
                     /note="chicken beta-globin 5'HS4 insulator core sequence"
     promoter        546..1045
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     regulatory      1059..1068
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1070..1117
                     /label=FRT mutant
                     /note="FRT mutant"
     primer_bind     complement(1162..1178)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(1186..1202)
                     /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)."
     promoter        complement(1210..1240)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(1255..1276)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(1564..2152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(2326..3183)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(3184..3288)
                     /label=AmpR promoter
     primer_bind     3762..3778
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3801..3848
                     /label=FRT
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     misc_feature    3865..3968
                     /label=Ad SA
                     /note="adenovirus major late transcript splice acceptor 
                     (Friedrich and Soriano, 1991)"
     primer_bind     4001..4017
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4031..4064
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     4139..4155
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4162..4286
                     /label=attR4
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4330..4632)
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(4977..5633)
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
     promoter        complement(5687..5717)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an 'up' mutation"
     protein_bind    complement(5743..5866)
                     /label=attR3
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5894..5910)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    5959..6183
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cgcgccgggc cctcgcgact cgaggaataa attcgagctc acggggacag ccccccccca
       61 aagcccccag ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg
      121 ggctccgctc cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc
      181 ccgggcacgg ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg
      241 agcctgcaga cacctggggg atacggggaa aaagcttgag ctcacgggga cagccccccc
      301 ccaaagcccc cagggatgta attacgtccc tcccccgcta gggggcagca gcgagccgcc
      361 cggggctccg ctccggtccg gcgctccccc cgcatccccg agccggcagc gtgcggggac
      421 agcccgggca cggggaaggt ggcacgggat cgctttcctc tgaacgcttc tcgctgctct
      481 ttgagcctgc agacacctgg gggatacggg gaaaaagctt gtcgacgaat aaattcaatt
      541 ctaccgggta ggggaggcgc ttttcccaag gcagtctgga gcatgcgctt tagcagcccc
      601 gctgggcact tggcgctaca caagtggcct ctggcctcgc acacattcca catccaccgg
      661 taggcgccaa ccggctccgt tctttggtgg ccccttcgcg ccaccttcta ctcctcccct
      721 agtcaggaag ttcccccccg ccccgcagct cgcgtcgtgc aggacgtgac aaatggaagt
      781 agcacgtctc actagtctcg tgcagatgga cagcaccgct gagcaatgga agcgggtagg
      841 cctttggggc agcggccaat agcagctttg ctccttcgct ttctgggctc agaggctggg
      901 aaggggtggg tccgggggcg ggctcagggg cgggctcagg ggcggggcgg gcgcccgaag
      961 gtcctccgga ggcccggcat tctgcacgct tcaaaagcgc acgtctgccg cgctgttctc
     1021 ctcttcctca tctccgggcc tttcgacctg caatcgccgc caccatggag aagttcctat
     1081 tccgaagttc ctattcttca aaaggtatag gaacttccga tcgggatccc caatcctgca
     1141 tgcaagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     1201 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     1261 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     1321 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     1381 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     1441 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     1501 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     1561 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     1621 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     1681 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     1741 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     1801 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     1861 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     1921 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     1981 gcctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt
     2041 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     2101 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     2161 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     2221 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     2281 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     2341 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     2401 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     2461 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     2521 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     2581 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     2641 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     2701 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     2761 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     2821 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     2881 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     2941 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     3001 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     3061 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     3121 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     3181 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
     3241 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     3301 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     3361 gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3421 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3481 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     3541 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     3601 gagaaaatac cgcatcaggc gccattcgcc attcaggctg cgcaactgtt gggaagggcg
     3661 atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg
     3721 attaagttgg gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga
     3781 attccacaaa agtgcgatcg gaagttccta ttccgaagtt cctattctct agaaagtata
     3841 ggaacttctt aattaaggga tctgtagggc gcagtagtcc agggtttcct tgatgatgtc
     3901 atacttatcc tgtccctttt ttttccacag ctcgcggttg aggacaaact cttcgcggtc
     3961 tttccagtgg ggatcgacgg tatcgtagag tcgaggccgc tctagaacta gtggatccgg
     4021 aacccttaat ataacttcgt ataatgtatg ctatacgaag ttattaggtc cctcgacctg
     4081 cagcccaagc tagcttatcg ataccgtcga cnctcgagnt tttcccagtc acgacgttgt
     4141 aaaacgacgg ccagtgaatt atcaactttg tatagaaaag ttgaacgaga aacgtaaaat
     4201 gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac ataatactgt
     4261 aaaacacaac atatccagtc actatggtcg acctgcagac tggctgtgta taagggagcc
     4321 tgacatttat attccccaga acatcaggtt aatggcgttt ttgatgtcat tttcgcggtg
     4381 gctgagatca gccacttctt ccccgataac ggagaccggc acactggcca tatcggtggt
     4441 catcatgcgc cagctttcat ccccgatatg caccaccggg taaagttcac gggggacttt
     4501 atctgacagc agacgtgcac tggccagggg gatcaccatc cgtcgcccgg gcgtgtcaat
     4561 aatatcactc tgtacatcca caaacagacg ataacggctc tctcttttat aggtgtaaac
     4621 cttaaactgc atttcaccag cccctgttct cgtcggcaaa agagccgttc atttcaataa
     4681 accgggcgac ctcagccatc ccttcctgat tttccgcttt ccagcgttcg gcacgcagac
     4741 gacgggcttc attctgcatg gttgtgctta ccgaaccgga gatattgaca tcatatatgc
     4801 cttgagcaac tgatagctgt cgctgtcaac tgtcactgta atacgctgct tcatagcata
     4861 cctctttttg acatacttcg ggtatacata tcagtatata ttcttatacc gcaaaaatca
     4921 gcgcgcaaat acgcatactg ttatctggct tttagtaagc cggatcctct agattacgcc
     4981 ccgccctgcc actcatcgca gtactgttgt aattcattaa gcattctgcc gacatggaag
     5041 ccatcacaaa cggcatgatg aacctgaatc gccagcggca tcagcacctt gtcgccttgc
     5101 gtataatatt tgcccatggt gaaaacgggg gcgaagaagt tgtccatatt ggccacgttt
     5161 aaatcaaaac tggtgaaact cacccaggga ttggctgaga cgaaaaacat attctcaata
     5221 aaccctttag ggaaataggc caggttttca ccgtaacacg ccacatcttg cgaatatatg
     5281 tgtagaaact gccggaaatc gtcgtggtat tcactccaga gcgatgaaaa cgtttcagtt
     5341 tgctcatgga aaacggtgta acaagggtga acactatccc atatcaccag ctcaccgtct
     5401 ttcattgcca tacggaattc cggatgagca ttcatcaggc gggcaagaat gtgaataaag
     5461 gccggataaa acttgtgctt atttttcttt acggtcttta aaaaggccgt aatatccagc
     5521 tgaacggtct ggttataggt acattgagca actgactgaa atgcctcaaa atgttcttta
     5581 cgatgccatt gggatatatc aacggtggta tatccagtga tttttttctc cattttagct
     5641 tccttagctc ctgaaaatct cgacggatcc taactcaaaa tccacacatt atacgagccg
     5701 gaagcataaa gtgtaaagcc tgggggtgcc taatgcggcc gccatagtga ctggatatgt
     5761 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     5821 tttatatcat tttacgtttc tcgttcaact ttattataca tagttgatag cttgatagct
     5881 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc gggccgcgag
     5941 ctcgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc
     6001 ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg
     6061 aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg
     6121 acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta
     6181 tggcttctga ggcggaaaga accagctggg gctcgatcct ctagttgg
//
