LOCUS       sequence-166106-        5571 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  hsDCX-pHAT N-terminal 6xHis-tag, C-terminal Strep-tagII.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5571)
  AUTHORS   Bechstedt S, Brouhard GJ
  TITLE     Doublecortin recognizes the 13-protofilament microtubule 
            cooperatively and tracks microtubule ends.
  JOURNAL   Dev Cell. 2012 Jul 17;23(1):181-92. doi: 
            10.1016/j.devcel.2012.05.006. Epub 2012 Jun 21.
  PUBMED    22727374
REFERENCE   2  (bases 1 to 5571)
  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"; doi: 
            "10.1016/j.devcel.2012.05.006"; journalName: "Dev Cell"; date: 
            "2012-07-17- 17"; volume: "23"; issue: "1"; pages: "181-92"
COMMENT     Sequence Label: hsDCX-pHAT
FEATURES             Location/Qualifiers
     source          1..5571
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    19..43
                     /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)."
     RBS             56..78
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             92..109
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             116..139
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             1277..1300
                     /codon_start=1
                     /product="peptide that binds Strep-Tactin(R), an engineered
                     form of streptavidin"
                     /label=Strep-Tag II
                     /translation="WSHPQFEK"
     promoter        complement(1316..1334)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1317..1334)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1392..1410)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1406..1453
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(1689..1710)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1860..2942)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPSTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     2904..2923
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2943..3020)
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     primer_bind     3166..3185
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     complement(3341..3360)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3460..3482
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3520..3538)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3606..3710
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3711..3779,3780..4571)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCHTLLSRIDAGQEQLGRRARYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4742..5330
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5231..5250
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5484..5501
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg aattgtgagc ggataacaat tcctctagca ataattttgt
       61 ttaactttaa gaaggagata tatccatgag tcatcaccat caccatcact ccatgctgga
      121 agttctgttc caggggccca ccatgggtcg acagctgagg ggagctatgg aacttgattt
      181 tggacacttt gacgaaagag ataagacatc caggaacatg cgaggctccc ggatgaatgg
      241 gttgcctagc cccactcaca gcgcccactg tagcttctac cgaaccagaa ccttgcaggc
      301 actgagtaat gagaagaaag ccaagaaggt acgtttctac cgcaatgggg accgctactt
      361 caaggggatt gtgtacgctg tgtcctctga ccgttttcgc agctttgacg ccttgctggc
      421 tgacctgacg cgatctctgt ccgacaacat caacctgcct cagggagtgc gttacattta
      481 caccattgat ggatccagga agatcggaag catggatgaa ctggaggaag gggaaagcta
      541 tgtctgttcc tcagacaact tctttaaaaa ggtggagtac accaagaatg tcaatcccaa
      601 ctggtctgtc aacgtaaaaa catctgccaa tatgaaagcc ccccagtctt tggctagcag
      661 caacagtgca caggccaggg agaacaagga ctttgtgcgc cccaagctgg ttaccatcat
      721 ccgcagtggg gtgaagcctc ggaaggctgt gcgtgtgctt ctgaacaaga agacagccca
      781 ctcttttgag caagtcctca ctgatatcac agaagccatc aaactggaga ccggggttgt
      841 caaaaaactc tacactctgg atggaaaaca ggtaacttgt ctccatgatt tctttggtga
      901 tgatgatgtg tttattgcct gtggtcctga aaaatttcgc tatgctcagg atgatttttc
      961 tctggatgaa aatgaatgcc gagtcatgaa gggaaaccca tcagccacag ctggcccaaa
     1021 ggcatcccca acacctcaga agacttcagc caagagccct ggtcctatgc gccgaagcaa
     1081 gtctccagct gactcaggta acgaccaaga cgcaaacgga acctccagca gccagctctc
     1141 tacccccaag tctaagcagt ctcccatctc tacgcccacc agtcctggca gcctccggaa
     1201 gcacaaggac ctgtacctgc ctctgtcctt ggatgactcg gactcgcttg gtgattccat
     1261 gagttcccct cagccctgga gccacccgca gttcgaaaaa tgaaagcttg agtattctat
     1321 agtgtcacct aaatcccagc ttgatccggc tgctaacaaa gcccgaaagg aagctgagtt
     1381 ggctgctgcc accgctgagc aataactagc ataacccctt ggggcctcta aacgggtctt
     1441 gaggggtttt ttgctgaaag gaggaactat atccggataa cctggcgtaa tagcgaagag
     1501 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gaaattgtaa
     1561 gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc
     1621 aataggccga aatcggcaaa atcccttata aatcaaaaga atagaccgag atagtgttga
     1681 gtgttgttcc agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag
     1741 ggcgaaaaac cgtctatcag ggcgatggcc cacagcttgc atgcctgcag gtcggaagca
     1801 taaagtgtaa agcctggggt gcctaatgag tgagctaact tacattaatt gcgttgcgct
     1861 cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac
     1921 gcgcggggag aggcggtttg cgtattgggc gccagggtgg tttttctttt caccagtgag
     1981 acgggcaaca gctgattgcc cttcaccgcc tggccctgag agagttgcag caagcggtcc
     2041 acgctggttt gccccagcag gcgaaaatcc tgtttgatgg tggttgacgg cgggatataa
     2101 catgagctgt cttcggtatc gtcgtatccc actaccgaga tatccgcacc aacgcgcagc
     2161 ccggactcgg taatggcgcg cattgcgccc agcgccatct gatcgttggc aaccagcatc
     2221 gcagtgggaa cgatgccctc attcagcatt tgcatggttt gttgaaaacc ggacatggca
     2281 ctccagtcgc cttcccgttc cgctatcggc tgaatttgat tgcgagtgag atatttatgc
     2341 cagccagcca gacgcagacg cgccgagaca gaacttaatg ggcccgctaa cagcgcgatt
     2401 tgctggtgac ccaatgcgac cagatgctcc acgcccagtc gcgtaccgtc ttcatgggag
     2461 aaaataatac tgttgatggg tgtctggtca gagacatcaa gaaataacgc cggaacatta
     2521 gtgcaggcag cttccacagc aatggcatcc tggtcatcca gcggatagtt aatgatcagc
     2581 ccactgacgc gttgcgcgag aagattgtgc accgccgctt tacaggcttc gacgccgctt
     2641 cgttctacca tcgacaccac cacgctggca cccagttgat cggcgcgaga tttaatcgcc
     2701 gcgacaattt gcgacggcgc gtgcagggcc agactggagg tggcaacgcc aatcagcaac
     2761 gactgtttgc ccgccagttg ttgtgccacg cggttgggaa tgtaattcag ctccgccatc
     2821 gccgcttcca ctttttcccg cgttttcgca gaaacgtggc tggcctggtt caccacgcgg
     2881 gaaacggtct gataagagac accggcatac tctgcgacat cgtataacgt tactggtttc
     2941 acattcacca ccctgaattg actctcttcc gggcgctatc atgccatacc gcgaaaggtt
     3001 ttgcaccatt cgatggtgtc gggtaccgag ctcgaattta cgtgaaccat cacccaaatc
     3061 aagttttttg cggtcgaggt gccgtaaagc tctaaatcgg aaccctaaag ggagcccccg
     3121 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     3181 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     3241 cgccgcgctt aatgcgccgc tacagggcgc gtcgcctgat gcggtatttt ctccttacgc
     3301 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
     3361 catagttaag ccagccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc
     3421 tgctcccggc atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga
     3481 ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg cctcgtgata cgcctatttt
     3541 tataggttaa tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa
     3601 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3661 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc
     3721 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     3781 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     3841 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     3901 ttccaatgat gagcactttt aaagttctgc tatgtcatac actattatcc cgtattgacg
     3961 ccgggcaaga gcaactcggt cgccgggcgc ggtattctca gaatgacttg gttgagtact
     4021 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     4081 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     4141 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     4201 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa
     4261 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     4321 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     4381 cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca
     4441 ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     4501 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     4561 agcattggta actgtcagac caagtttact catatatact ttagattgat ttaaaacttc
     4621 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     4681 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     4741 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     4801 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     4861 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     4921 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     4981 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     5041 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     5101 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     5161 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     5221 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     5281 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     5341 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     5401 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     5461 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa
     5521 tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctga t
//
