LOCUS       sequence-362060-        6668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KIF1A(1-393)(P305L) fused with leucine zipper and 
            His tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6668)
  AUTHORS   Anazawa Y, Kita T, Iguchi R, Hayashi K, Niwa S
  TITLE     De novo mutations in KIF1A-associated neuronal disorder (KAND) 
            dominant-negatively inhibit motor activity and axonal transport of 
            synaptic vesicle precursors.
  JOURNAL   Proc Natl Acad Sci U S A. 2022 Aug 9;119(32):e2113795119. doi: 
            10.1073/pnas.2113795119. Epub 2022 Aug 2.
  PUBMED    35917346
REFERENCE   2  (bases 1 to 6668)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2022-08-9"; volume: "119(32):e2113795119. 
            doi"; pages: " 10.1073/pnas.2113795119. Epub 2022 Aug 2"
COMMENT     Sequence Label: pSN687
FEATURES             Location/Qualifiers
     source          1..6668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(45..63)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      59..106
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      143..598
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(230..249)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     440..461
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        624..728
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(729..797,798..1589)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(947..966)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1760..2348
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2249..2268
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2502..2519
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2534..2673
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2599..2618)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2759..2781
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2775..2966)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3775..4857)
                     /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
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     4819..4838
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4858..4935)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5141..5160
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5244..5263
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5244..5262
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5263..5287
                     /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             5302..5324
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6643..6660
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
ORIGIN
        1 ggctgctaac aaagcccgaa aggaagctga gttggctgct gccaccgctg agcaataact
       61 agcataaccc cttggggcct ctaaacgggt cttgaggggt tttttgctga aaggaggaac
      121 tatatccgga ttggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
      181 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
      241 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
      301 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
      361 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
      421 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
      481 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
      541 gagctgattt aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag
      601 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
      661 caaatatgta tccgctcatg tgacaataac cctgataaat gcttcaataa tattgaaaaa
      721 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt
      781 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
      841 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
      901 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
      961 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     1021 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     1081 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     1141 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     1201 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     1261 ccacgatgcc tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     1321 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     1381 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     1441 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     1501 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     1561 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     1621 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     1681 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     1741 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     1801 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     1861 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc
     1921 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1981 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     2041 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     2101 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     2161 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     2221 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     2281 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     2341 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     2401 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     2461 agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     2521 aagcggaaga gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc
     2581 gcaatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact
     2641 ccgctatcgc tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac
     2701 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     2761 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg
     2821 taaagctcat cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc
     2881 agctcgttga gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta
     2941 agggcggttt tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg
     3001 ggggtaatga taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa
     3061 catgcccggt tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac
     3121 cagagaaaaa tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca
     3181 cagggtagcc agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac
     3241 ttccgcgttt ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag
     3301 gtcgcagacg ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc
     3361 tgctaaccag taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc
     3421 atgcgcaccc gtggggccgc catgccggcg ataatggcct gcttctcgcc gaaacgtttg
     3481 gtggcgggac cagtgacgaa ggcttgagcg agggcgtgca agattccgaa taccgcaagc
     3541 gacaggccga tcatcgtcgc gctccagcga aagcggtcct cgccgaaaat gacccagagc
     3601 gctgccggca cctgtcctac gagttgcatg ataaagaaga cagtcataag tgcggcgacg
     3661 atagtcatgc cccgcgccca ccggaaggag ctgactgggt tgaaggctct caagggcatc
     3721 ggtcgagatc ccggtgccta atgagtgagc taacttacat taattgcgtt gcgctcactg
     3781 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     3841 gggagaggcg gtttgcgtat tgggcgccag ggtggttttt cttttcacca gtgagacggg
     3901 caacagctga ttgcccttca ccgcctggcc ctgagagagt tgcagcaagc ggtccacgct
     3961 ggtttgcccc agcaggcgaa aatcctgttt gatggtggtt aacggcggga tataacatga
     4021 gctgtcttcg gtatcgtcgt atcccactac cgagatatcc gcaccaacgc gcagcccgga
     4081 ctcggtaatg gcgcgcattg cgcccagcgc catctgatcg ttggcaacca gcatcgcagt
     4141 gggaacgatg ccctcattca gcatttgcat ggtttgttga aaaccggaca tggcactcca
     4201 gtcgccttcc cgttccgcta tcggctgaat ttgattgcga gtgagatatt tatgccagcc
     4261 agccagacgc agacgcgccg agacagaact taatgggccc gctaacagcg cgatttgctg
     4321 gtgacccaat gcgaccagat gctccacgcc cagtcgcgta ccgtcttcat gggagaaaat
     4381 aatactgttg atgggtgtct ggtcagagac atcaagaaat aacgccggaa cattagtgca
     4441 ggcagcttcc acagcaatgg catcctggtc atccagcgga tagttaatga tcagcccact
     4501 gacgcgttgc gcgagaagat tgtgcaccgc cgctttacag gcttcgacgc cgcttcgttc
     4561 taccatcgac accaccacgc tggcacccag ttgatcggcg cgagatttaa tcgccgcgac
     4621 aatttgcgac ggcgcgtgca gggccagact ggaggtggca acgccaatca gcaacgactg
     4681 tttgcccgcc agttgttgtg ccacgcggtt gggaatgtaa ttcagctccg ccatcgccgc
     4741 ttccactttt tcccgcgttt tcgcagaaac gtggctggcc tggttcacca cgcgggaaac
     4801 ggtctgataa gagacaccgg catactctgc gacatcgtat aacgttactg gtttcacatt
     4861 caccaccctg aattgactct cttccgggcg ctatcatgcc ataccgcgaa aggttttgcg
     4921 ccattcgatg gtgtccggga tctcgacgct ctcccttatg cgactcctgc attaggaagc
     4981 agcccagtag taggttgagg ccgttgagca ccgccgccgc aaggaatggt gcatgcaagg
     5041 agatggcgcc caacagtccc ccggccacgg ggcctgccac catacccacg ccgaaacaag
     5101 cgctcatgag cccgaagtgg cgagcccgat cttccccatc ggtgatgtcg gcgatatagg
     5161 cgccagcaac cgcacctgtg gcgccggtga tgccggccac gatgcgtccg gcgtagagga
     5221 tcgagatctc gatcccgcga aattaatacg actcactata ggggaattgt gagcggataa
     5281 caattcccct ctagaaataa ttttgtttaa ctttaagaag gagatataca tatgggcagc
     5341 agcatggccg gggcttcggt gaaggtggcg gtgcgggtcc gccccttcaa ttcccgggaa
     5401 atgagccgtg actccaagtg catcattcag atgtctggaa gcaccaccac cattgttaac
     5461 cccaaacagc ccaaggagac gcccaaaagc ttcagctttg actactccta ctggtcgcac
     5521 acctcacctg aggacatcaa ctacgcgtcg cagaagcagg tgtaccggga catcggcgag
     5581 gagatgctgc agcatgcctt tgagggatac aacgtgtgca tcttcgccta tgggcagacg
     5641 ggtgccggca agtcctacac catgatgggc aagcaggaga aggaccagca gggcatcatc
     5701 ccacagctct gcgaggacct cttctctcgg atcaacgaca cgaccaacga caacatgtcc
     5761 tactccgtgg aggtcagcta catggagatt tactgtgagc gcgtccgtga cctcctgaac
     5821 cccaagaaca agggcaacct tcgcgtgagg gagcacccac tgctggggcc ctacgtggag
     5881 gacctctcca agctggctgt cacctcctac aatgacatcc aggacctcat ggactcaggg
     5941 aacaaggcca ggaccgtggc ggccaccaac atgaatgaga ccagcagtcg ctcccacgcc
     6001 gtcttcaaca tcatcttcac ccagaagcgc catgacgcag agaccaatat caccacggag
     6061 aaggtgagca aaatcagcct ggtggacctg gctgggagcg agcgggctga ctccacggga
     6121 gccaagggca cgcgcctcaa ggagggggcc aacatcaaca agtcgctgac caccctgggc
     6181 aaggtcatct ccgccctggc tgaaatggac tccggaccca acaagaacaa gaaaaagaag
     6241 aagacagatt tcattctgta ccgagattcc gtgttgacct ggctcctccg ggaaaacctg
     6301 ggcggtaact caaggacagc tatggtggca gccttgagtc ctgcagacat caactacgat
     6361 gagaccctta gcacgctgag gtatgctgac cgggccaagc agatccgctg caatgctgtc
     6421 atcaatgagg accccaacaa caagctgatc cgcgagctga aggatgaggt gacccggctg
     6481 cgggaccttc tgtacgccca gggtcttggc gacatcactg acggcgcggg agtgaaacaa
     6541 cttgaagaca aggttgaaga attggcttcg aaaaattatc acttggaaaa tgaggttgcc
     6601 agattaaaga aattagtttg cggcagcggt agcgcactcg agcaccacca ccaccaccac
     6661 tgagatcc
//
