LOCUS       sequence-355088-        6668 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses human KIF1A(1-393) 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: pSN672
FEATURES             Location/Qualifiers
     source          1..6668
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(74..93)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     284..305
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        468..572
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(573..641,642..1433)
                     /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(791..810)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1604..2192
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2093..2112
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2346..2363
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2378..2517
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2443..2462)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2603..2625
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2619..2810)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(3619..4701)
                     /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     4663..4682
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4702..4779)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4985..5004
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     5088..5107
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        5088..5106
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    5107..5131
                     /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             5146..5168
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             6487..6504
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6557..6575)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      6571..6618
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      join(6655..6668,1..442)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
       61 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
      121 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
      181 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
      241 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
      301 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
      361 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
      421 aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc ggaaccccta
      481 tttgtttatt tttctaaata cattcaaata tgtatccgct catgtgacaa taaccctgat
      541 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
      601 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
      661 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
      721 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
      781 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
      841 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
      901 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
      961 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     1021 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     1081 ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca acgttgcgca
     1141 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     1201 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     1261 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     1321 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     1381 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     1441 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     1501 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     1561 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     1621 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     1681 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     1741 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     1801 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     1861 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     1921 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     1981 acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     2041 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     2101 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     2161 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     2221 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     2281 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     2341 agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct gatgcggtat tttctcctta
     2401 cgcatctgtg cggtatttca caccgcaatg gtgcactctc agtacaatct gctctgatgc
     2461 cgcatagtta agccagtata cactccgcta tcgctacgtg actgggtcat ggctgcgccc
     2521 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     2581 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     2641 ccgaaacgcg cgaggcagct gcggtaaagc tcatcagcgt ggtcgtgaag cgattcacag
     2701 atgtctgcct gttcatccgc gtccagctcg ttgagtttct ccagaagcgt taatgtctgg
     2761 cttctgataa agcgggccat gttaagggcg gttttttcct gtttggtcac tgatgcctcc
     2821 gtgtaagggg gatttctgtt catgggggta atgataccga tgaaacgaga gaggatgctc
     2881 acgatacggg ttactgatga tgaacatgcc cggttactgg aacgttgtga gggtaaacaa
     2941 ctggcggtat ggatgcggcg ggaccagaga aaaatcactc agggtcaatg ccagcgcttc
     3001 gttaatacag atgtaggtgt tccacagggt agccagcagc atcctgcgat gcagatccgg
     3061 aacataatgg tgcagggcgc tgacttccgc gtttccagac tttacgaaac acggaaaccg
     3121 aagaccattc atgttgttgc tcaggtcgca gacgttttgc agcagcagtc gcttcacgtt
     3181 cgctcgcgta tcggtgattc attctgctaa ccagtaaggc aaccccgcca gcctagccgg
     3241 gtcctcaacg acaggagcac gatcatgcgc acccgtgggg ccgccatgcc ggcgataatg
     3301 gcctgcttct cgccgaaacg tttggtggcg ggaccagtga cgaaggcttg agcgagggcg
     3361 tgcaagattc cgaataccgc aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg
     3421 tcctcgccga aaatgaccca gagcgctgcc ggcacctgtc ctacgagttg catgataaag
     3481 aagacagtca taagtgcggc gacgatagtc atgccccgcg cccaccggaa ggagctgact
     3541 gggttgaagg ctctcaaggg catcggtcga gatcccggtg cctaatgagt gagctaactt
     3601 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3661 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ccagggtggt
     3721 ttttcttttc accagtgaga cgggcaacag ctgattgccc ttcaccgcct ggccctgaga
     3781 gagttgcagc aagcggtcca cgctggtttg ccccagcagg cgaaaatcct gtttgatggt
     3841 ggttaacggc gggatataac atgagctgtc ttcggtatcg tcgtatccca ctaccgagat
     3901 atccgcacca acgcgcagcc cggactcggt aatggcgcgc attgcgccca gcgccatctg
     3961 atcgttggca accagcatcg cagtgggaac gatgccctca ttcagcattt gcatggtttg
     4021 ttgaaaaccg gacatggcac tccagtcgcc ttcccgttcc gctatcggct gaatttgatt
     4081 gcgagtgaga tatttatgcc agccagccag acgcagacgc gccgagacag aacttaatgg
     4141 gcccgctaac agcgcgattt gctggtgacc caatgcgacc agatgctcca cgcccagtcg
     4201 cgtaccgtct tcatgggaga aaataatact gttgatgggt gtctggtcag agacatcaag
     4261 aaataacgcc ggaacattag tgcaggcagc ttccacagca atggcatcct ggtcatccag
     4321 cggatagtta atgatcagcc cactgacgcg ttgcgcgaga agattgtgca ccgccgcttt
     4381 acaggcttcg acgccgcttc gttctaccat cgacaccacc acgctggcac ccagttgatc
     4441 ggcgcgagat ttaatcgccg cgacaatttg cgacggcgcg tgcagggcca gactggaggt
     4501 ggcaacgcca atcagcaacg actgtttgcc cgccagttgt tgtgccacgc ggttgggaat
     4561 gtaattcagc tccgccatcg ccgcttccac tttttcccgc gttttcgcag aaacgtggct
     4621 ggcctggttc accacgcggg aaacggtctg ataagagaca ccggcatact ctgcgacatc
     4681 gtataacgtt actggtttca cattcaccac cctgaattga ctctcttccg ggcgctatca
     4741 tgccataccg cgaaaggttt tgcgccattc gatggtgtcc gggatctcga cgctctccct
     4801 tatgcgactc ctgcattagg aagcagccca gtagtaggtt gaggccgttg agcaccgccg
     4861 ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag tcccccggcc acggggcctg
     4921 ccaccatacc cacgccgaaa caagcgctca tgagcccgaa gtggcgagcc cgatcttccc
     4981 catcggtgat gtcggcgata taggcgccag caaccgcacc tgtggcgccg gtgatgccgg
     5041 ccacgatgcg tccggcgtag aggatcgaga tctcgatccc gcgaaattaa tacgactcac
     5101 tataggggaa ttgtgagcgg ataacaattc ccctctagaa ataattttgt ttaactttaa
     5161 gaaggagata tacatatggg cagcagcatg gccggggctt cggtgaaggt ggcggtgcgg
     5221 gtccgcccct tcaattcccg ggaaatgagc cgtgactcca agtgcatcat tcagatgtct
     5281 ggaagcacca ccaccattgt taaccccaaa cagcccaagg agacgcccaa aagcttcagc
     5341 tttgactact cctactggtc gcacacctca cctgaggaca tcaactacgc gtcgcagaag
     5401 caggtgtacc gggacatcgg cgaggagatg ctgcagcatg cctttgaggg atacaacgtg
     5461 tgcatcttcg cctatgggca gacgggtgcc ggcaagtcct acaccatgat gggcaagcag
     5521 gagaaggacc agcagggcat catcccacag ctctgcgagg acctcttctc tcggatcaac
     5581 gacacgacca acgacaacat gtcctactcc gtggaggtca gctacatgga gatttactgt
     5641 gagcgcgtcc gtgacctcct gaaccccaag aacaagggca accttcgcgt gagggagcac
     5701 ccactgctgg ggccctacgt ggaggacctc tccaagctgg ctgtcacctc ctacaatgac
     5761 atccaggacc tcatggactc agggaacaag gccaggaccg tggcggccac caacatgaat
     5821 gagaccagca gtcgctccca cgccgtcttc aacatcatct tcacccagaa gcgccatgac
     5881 gcagagacca atatcaccac ggagaaggtg agcaaaatca gcctggtgga cctggctggg
     5941 agcgagcggg ctgactccac gggagccaag ggcacgcgcc tcaaggaggg ggccaacatc
     6001 aacaagtcgc tgaccaccct gggcaaggtc atctccgccc tggctgaaat ggactccgga
     6061 cccaacaaga acaagaaaaa gaagaagaca gatttcattc cgtaccgaga ttccgtgttg
     6121 acctggctcc tccgggaaaa cctgggcggt aactcaagga cagctatggt ggcagccttg
     6181 agtcctgcag acatcaacta cgatgagacc cttagcacgc tgaggtatgc tgaccgggcc
     6241 aagcagatcc gctgcaatgc tgtcatcaat gaggacccca acaacaagct gatccgcgag
     6301 ctgaaggatg aggtgacccg gctgcgggac cttctgtacg cccagggtct tggcgacatc
     6361 actgacggcg cgggagtgaa acaacttgaa gacaaggttg aagaattggc ttcgaaaaat
     6421 tatcacttgg aaaatgaggt tgccagatta aagaaattag tttgcggcag cggtagcgca
     6481 ctcgagcacc accaccacca ccactgagat ccggctgcta acaaagcccg aaaggaagct
     6541 gagttggctg ctgccaccgc tgagcaataa ctagcataac cccttggggc ctctaaacgg
     6601 gtcttgaggg gttttttgct gaaaggagga actatatccg gattggcgaa tgggacgcgc
     6661 cctgtagc
//
