LOCUS       40924_570        7385 bp DNA     circular SYN 17-DEC-2018
DEFINITION  Cloning vector DBH-FLPo, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7385)
  AUTHORS   Sun JJ, Ray R.
  TITLE     Generation of Two Noradrenergic-Specific 
            Dopamine-Beta-Hydroxylase-FLPo Knock-In Mice Using 
            CRISPR/Cas9-Mediated Targeting in Embryonic Stem Cells
  JOURNAL   PLoS ONE 11 (7), E0159474 (2016)
  PUBMED    27441631
REFERENCE   2  (bases 1 to 7385)
  AUTHORS   Sun JJ, Ray R.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-APR-2016) Neuroscience, Baylor College of Medicine, 
            One Baylor Plaza Room T707, Houston, TX 77030, USA
REFERENCE   3  (bases 1 to 7385)
  TITLE     Direct Submission
REFERENCE   4  (bases 1 to 7385)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "PLoS ONE"; 
            date: "2016"; volume: "11"; issue: "7"; pages: "E0159474"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (27-APR-2016) Neuroscience, Baylor College of Medicine, One Baylor 
            Plaza Room T707, Houston, TX 77030, USA"
COMMENT     SGRef: number: 3; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7385
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      18..562
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     misc_feature    842..1838
                     /label=5' homology arm
                     /note="5' homology arm"
     CDS             1842..3137
                     /label=FLPo
                     /note="nuclear-targeted site-specific recombinase"
     polyA_signal    3171..3395
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3403..3436
                     /label=lox2272
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     promoter        3443..3942
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     promoter        3954..4001
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter"
     CDS             4020..4820
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     regulatory      4837..5139
                     /label=bghpA
                     /note="bghpA"
                     /regulatory_class="other"
     polyA_signal    4861..5085
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     misc_feature    5140..5173
                     /label=Lox2722
                     /note="Lox2722"
     protein_bind    complement(5140..5173)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    5198..6197
                     /label=3' homology arm with deletion
                     /note="3' homology arm with deletion"
     promoter        6273..6377
                     /label=AmpR promoter
     CDS             6378..7235
                     /label=AmpR
                     /note="beta-lactamase"
ORIGIN
        1 ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg ctctgaaaac
       61 gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa ctctttgaac
      121 cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt ttagccttaa
      181 ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg ctgccagtgg
      241 tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata aggcgcagcg
      301 gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg cctacccgga
      361 actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg acaccggtaa
      421 accgaaaggc aggaacagga gagcgcacga gggagccgcc aggggaaacg cctggtatct
      481 ttatagtcct gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc
      541 aggggggcgg agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt
      601 atcttcctgg catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc
      661 agtcgaacga ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac
      721 atattctgct gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga
      781 caccctcatc agtgccaaca tagtaagcca gtatacactc cgctagcgaa ttgataggcc
      841 tagaggctca ggctgggcta ctgtgtttct gagaaagaag caggaagaaa aagagggaga
      901 gaaggaagga gagaaggact cacaactagg ttggagggac ctatagtcac agtgaggtac
      961 agaggtggat ggctttggat acaagcagca gaccaagtag aatttcctgg tgttggaggt
     1021 atggacagta ctggttctca tcatgagaca gacagggctg agcaatagga aggccaacct
     1081 gtgcaggggc cctatggcag gtttgggagg gacagcgctg ggtacagctg acctgagaag
     1141 caagaggaag ccctgggttt gagagcttca gggagatggg aacaggtatg tcccccaccc
     1201 tcctttcccc taagtactgt ccctacagcc acgagagccc attcacccct ttctgcctgc
     1261 cctgctgtct ctgcacccac tccttctgta gcagccgtgg cgccagctgg agcccgtggg
     1321 gaaacaggag aaaaagtggg aacagccggc cttggagaga gtgtctaaac acggactatt
     1381 tggggactgc cttttatttt tagtttgaaa cttatccacc ccccagtatt gttaaaaaca
     1441 agagccagga acccaccact cactgtcact caggaaagag ccccttagac ctcagttgga
     1501 gcaggtggag aggacctggt atctggctgc ctatgaggcc ctccgcaggg cgtggctgga
     1561 ggtgtcctta ggacctatgt ctgcagagag tagctgtttc caacaggcgt cagagatttg
     1621 ttggaggaca tcagccactc tgcttcgatt ctcctgaaga tgtccatgtg tcattagtgc
     1681 caattagggg agatgtgttt gccccagggc atggactggt gggagggcca ccaagacaat
     1741 tcaattcccc accagacaaa tgtgattagg tgcagcctgg cccaacccca ctggacaggc
     1801 ataaatggca gagtggggtt ggggtgctca tccctgccac catggctcct aagaagaaga
     1861 ggaaggtgat gagccagttc gacatcctgt gcaagacccc ccccaaggtg ctggtgcggc
     1921 agttcgtgga gagattcgag aggcccagcg gcgagaagat cgccagctgt gccgccgagc
     1981 tgacctacct gtgctggatg atcacccaca acggcaccgc catcaagagg gccaccttca
     2041 tgagctacaa caccatcatc agcaacagcc tgagcttcga catcgtgaac aagagcctgc
     2101 agttcaagta caagacccag aaggccacca tcctggaggc cagcctgaag aagctgatcc
     2161 ccgcctggga gttcaccatc atcccttaca acggccagaa gcaccagagc gacatcaccg
     2221 acatcgtgtc cagcctgcag ctgcagttcg agagcagcga ggaggccgac aagggcaaca
     2281 gccacagcaa gaagatgctg aaggccctgc tgtccgaggg cgagagcatc tgggagatca
     2341 ccgagaagat cctgaacagc ttcgagtaca ccagcaggtt caccaagacc aagaccctgt
     2401 accagttcct gttcctggcc acattcatca actgcggcag gttcagcgac atcaagaacg
     2461 tggaccccaa gagcttcaag ctggtgcaga acaagtacct gggcgtgatc attcagtgcc
     2521 tggtgaccga gaccaagaca agcgtgtcca ggcacatcta ctttttcagc gccagaggca
     2581 ggatcgaccc cctggtgtac ctggacgagt tcctgaggaa cagcgagccc gtgctgaaga
     2641 gagtgaacag gaccggcaac agcagcagca acaagcagga gtaccagctg ctgaaggaca
     2701 acctggtgcg cagctacaac aaggccctga agaagaacgc cccctacccc atcttcgcta
     2761 tcaagaacgg ccctaagagc cacatcggca ggcacctgat gaccagcttt ctgagcatga
     2821 agggcctgac cgagctgaca aacgtggtgg gcaactggag cgacaagagg gcctccgccg
     2881 tggccaggac cacctacacc caccagatca ccgccatccc cgaccactac ttcgccctgg
     2941 tgtccaggta ctacgcctac gaccccatca gcaaggagat gatcgccctg aaggacgaga
     3001 ccaaccccat cgaggagtgg cagcacatcg agcagctgaa gggcagcgcc gagggcagca
     3061 tcagataccc cgcctggaac ggcatcatca gccaggaggt gctggactac ctgagcagct
     3121 acatcaacag gcggatctga tgagatctag agctcgctga tcagcctcga ctgtgccttc
     3181 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     3241 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     3301 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     3361 tagcaggcat gctggggatg cggtgggctc tatggatcat ttataacttc gtataaagta
     3421 tcctatacga agttattcta ccgggtaggg gaggcgcttt tcccaaggca gtctggagca
     3481 tgcgctttag cagccccgct gggcacttgg cgctacacaa gtggcctctg gcctcgcaca
     3541 cattccacat ccaccggtag gcgccaaccg gctccgttct ttggtggccc cttcgcgcca
     3601 ccttctactc ctcccctagt caggaagttc ccccccgccc cgcagctcgc gtcgtgcagg
     3661 acgtgacaaa tggaagtagc acgtctcact agtctcgtgc agatggacag caccgctgag
     3721 caatggaagc gggtaggcct ttggggcagc ggccaatagc agctttgctc cttcgctttc
     3781 tgggctcaga ggctgggaag gggtgggtcc gggggcgggc tcaggggcgg gctcaggggc
     3841 ggggcgggcg cccgaaggtc ctccggaggc ccggcattct gcacgcttca aaagcgcacg
     3901 tctgccgcgc tgttctcctc ttcctcatct ccgggccttt cgacctgcag cctgttgaca
     3961 attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg aactaaacca
     4021 tgggatcggc cattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4081 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4141 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4201 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4261 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4321 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4381 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4441 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4501 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4561 tgcccgacgg cgatgatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4621 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4681 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4741 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4801 gccttcttga cgagttcttc tgaggggatc aattctctag agctcgctga tcagcctcga
     4861 ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc
     4921 tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc
     4981 tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt
     5041 gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa
     5101 gaaccagctg gggctcgact agagcttgcg gaacccttaa taacttcgta taaagtatcc
     5161 tatacgaagt tataaataat ttctagaaag cttaattggt ggctgcactg cggggctcgg
     5221 agcctccaga gagccccttc ccctaccaca tccccctgga ccccgaaggg attttagagc
     5281 tctcatggaa cgtcagctat gtccaggaga tcatccactt ccagctccag gtgcaagggc
     5341 tgagggctgg ggtcctgttt ggaatgtcag atcgaggaga gatggagaac gcagatctca
     5401 tcatgctctg gactgatggg gacagggcct actttgcggt gagtcctctg ccatctttct
     5461 gtcttgtcct cacgtccata ccccccccac acacacacac acaagctggc tagcttcata
     5521 cagccctgct ctacagtcag caaagaaagc ttgcctactg cttctgtgtt tgatgtcact
     5581 aatctgaatt aagcccttca gcgtcataag gacactcact agagaatgga acagtaagta
     5641 gtctctcctt gaaatccttg actacacccc caaataatgg ttgcagcaat cctaaggaca
     5701 gtaagggcag atggaggacc cacatgccat gagtccccac ccacccacac acacatgagt
     5761 agagaccctg catatcctcc agttttacaa atagcaactc catgccacac taccagtaca
     5821 tgtgcacctg ttgtgggcca ggtggcatat ggctgaatag agcaggcagt atgcacacca
     5881 gcttgggaca cagaaacacg ccttctgtgc tatactcaat gatggtaaag gtagacacag
     5941 ataccactgc aacccgggtg tgggacaacc ccatcctcac cacacaagct gagcccaggc
     6001 caacagaatg cctctggttc cttctgttca aaagctacct gaacatctgc atgctagggt
     6061 agcaaacagg aaatctccta cagaccggag catccttcag ggcccctaca agctacaggg
     6121 atcagtagac ggaggataga tacccacaca tatcctctgt ggttctactc tttgtcctcc
     6181 tacctctaca gagggatatc atttaaataa tttctagaaa gcttaattcg taccgatcta
     6241 acgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     6301 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     6361 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     6421 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     6481 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     6541 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     6601 gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc cgcatacact
     6661 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     6721 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     6781 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     6841 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     6901 agcgtgacac cacgatgcct gcagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     6961 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     7021 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     7081 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     7141 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     7201 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     7261 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     7321 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     7381 acccc
//
