LOCUS       sequence-198298-        5422 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Generates shRNA specific to Rat KIF1A along with cell fill 
            pmTurqoise.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5422)
  AUTHORS   McVicker DP, Awe AM, Richters KE, Wilson RL, Cowdrey DA, Hu X, 
            Chapman ER, Dent EW
  TITLE     Transport of a kinesin-cargo pair along microtubules into dendritic 
            spines undergoing synaptic plasticity.
  JOURNAL   Nat Commun. 2016 Sep 23;7:12741. doi: 10.1038/ncomms12741.
  PUBMED    27658622
REFERENCE   2  (bases 1 to 5422)
  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: "Nat 
            Commun."; date: "2016-09-23"; volume: "7:12741. doi"; pages: " 
            10.1038/ncomms12741"
COMMENT     Sequence Label: pmTurqoise KIF1A shRNA
FEATURES             Location/Qualifiers
     source          1..5422
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..215
                     /label=H1 promoter
                     /note="human H1 RNA promoter"
     primer_bind     136..155
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     primer_bind     complement(284..300)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(285..301)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     promoter        complement(329..347)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(368..384)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(368..384)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(381..403)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    392..408
                     /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)."
     promoter        complement(join(416..422,423..440,441..446))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    461..482
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(599..616)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(770..1358)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(850..869)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1529..2320,2321..2389))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2152..2171
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2390..2494)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(2520..2975)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2657..2678)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2869..2888
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3102..3124
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3116..3133
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3117..3133
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3140..3159
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3140..3158
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     3191..3207
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3191..3207
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     polyA_signal    3245..3293
                     /note="synthetic polyadenylation signal"
     CDS             complement(3357..4151)
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3468..3487)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4078..4097
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     CDS             complement(join(4170..4883,4884..4886,4887..4889))
                     /codon_start=1
                     /product="enhanced monomeric variant of CFP (Goedhart et 
                     al., 2012)"
                     /label=mTurquoise2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYFSDNVYITADKQKNGIK
                     ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4215..4236)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4564..4583
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4823..4844
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     promoter        complement(4909..5412)
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     primer_bind     complement(4957..4976)
                     /label=mPGK-F
                     /note="Mouse PGK promoter, forward primer"
     primer_bind     5354..5377
                     /label=MSCV-rev
                     /note="Murine stem cell virus, reverse primer"
ORIGIN
        1 gaacgctgac gtcatcaacc cgctccaagg aatcgcgggc ccagtgtcac taggcgggaa
       61 cacccagcgc gcgtgcgccc tggcaggaag atggctgtga gggacagggg agtggcgccc
      121 tgcaatattt gcatgtcgct atgtgttctg ggaaatcacc ataaacgtga aatgtctttg
      181 gatttgggaa tcttataagt tctgtatgag accacagatc ccctacctat gtgaacggca
      241 agttcaagag acttgccgtt cacataggta tttttaagct tatcgatacc gtcgacctcg
      301 agggggggcc cggtaccagc ttttgttccc tttagtgagg gttaatttcg agcttggcgt
      361 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
      421 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
      481 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
      541 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
      601 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
      661 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
      721 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
      781 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
      841 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
      901 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
      961 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     1021 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     1081 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     1141 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     1201 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     1261 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     1321 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     1381 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     1441 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     1501 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     1561 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     1621 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     1681 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     1741 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     1801 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     1861 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     1921 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     1981 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     2041 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     2101 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     2161 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     2221 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     2281 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     2341 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     2401 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     2461 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     2521 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     2581 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     2641 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     2701 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     2761 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     2821 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     2881 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     2941 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     3001 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg
     3061 atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa
     3121 aacgacggcc agtgaattgt aatacgactc actatagggc gaattggagc tccaccgcgg
     3181 tggcggccgc tctagaacta gtggatcccc cgggctgcat ccccgatcct tatcgctatc
     3241 gattcacaca aaaaaccaac acacagatgt aatgaaaata aagatatttt attgcggcca
     3301 tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag tcccgctcag
     3361 aagaactcgt caagaaggcg atagaaggcg atgcgctgcg aatcgggagc ggcgataccg
     3421 taaagcacga ggaagcggtc agcccattcg ccgccaagct cttcagcaat atcacgggta
     3481 gccaacgcta tgtcctgata gcggtccgcc acacccagcc ggccacagtc gatgaatcca
     3541 gaaaagcggc cattttccac catgatattc ggcaagcagg catcgccatg ggtcacgacg
     3601 agatcctcgc cgtcgggcat gcgcgccttg agcctggcga acagttcggc tggcgcgagc
     3661 ccctgatgct cttcgtccag atcatcctga tcgacaagac cggcttccat ccgagtacgt
     3721 gctcgctcga tgcgatgttt cgcttggtgg tcgaatgggc aggtagccgg atcaagcgta
     3781 tgcagccgcc gcattgcatc agccatgatg gatactttct cggcaggagc aaggtgagat
     3841 gacaggagat cctgccccgg cacttcgccc aatagcagcc agtcccttcc cgcttcagtg
     3901 acaacgtcga gcacagctgc gcaaggaacg cccgtcgtgg ccagccacga tagccgcgct
     3961 gcctcgtcct gcagttcatt cagggcaccg gacaggtcgg tcttgacaaa aagaaccggg
     4021 cgcccctgcg ctgacagccg gaacacggcg gcatcagagc agccgattgt ctgttgtgcc
     4081 cagtcatagc cgaatagcct ctccacccaa gcggccggag aacctgcgtg caatccatct
     4141 tgttcaatca tggtggctct agccttaagt cacttgtaca gctcgtccat gccgagagtg
     4201 atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc gttggggtct
     4261 ttgctcagct tggactgggt gctcaggtag tggttgtcgg gcagcagcac ggggccgtcg
     4321 ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgccgccgtc ctcgatgttg
     4381 tggcggatct tgaagttggc cttgatgccg ttcttctgct tgtcggcggt gatatagacg
     4441 ttgtcgctaa agtagttgta ctccagcttg tgccccagga tgttgccgtc ctccttgaag
     4501 tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt cacctcggcg
     4561 cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac gtagccttcg
     4621 ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg ggcgaagcac
     4681 tgcacgcccc aggacagggt ggtcacgagg gtgggccagg gcacgggcag cttgccggtg
     4741 gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc ctcgccggac
     4801 acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg caccaccccg
     4861 gtgaacagct cctcgccctt gctcaccatg ctagcttggg ctgcaggtcg aaaggcccgg
     4921 agatgaggaa gaggagaaca gcgcggcaga cgtgcgcttt tgaagcgtgc agaatgccgg
     4981 gcctccggag gaccttcggg cgcccgcccc gcccctgagc ccgcccctga gcccgccccc
     5041 ggacccaccc cttcccagcc tctgagccca gaaagcgaag gagcaaagct gctattggcc
     5101 gctgccccaa aggcctaccc gcttccattg ctcagcggtg ctgtccatct gcacgagact
     5161 agtgagacgt gctacttcca tttgtcacgt cctgcacgac gcgagctgcg gggcgggggg
     5221 gaacttcctg actaggggag gagtagaagg tggcgcgaag gggccaccaa agaacggagc
     5281 cggttggcgc ctaccggccg gtggatgtgg aatgtgtgcg aggccagagg ccacttgtgt
     5341 agcgccaagt gcccagcggg gctgctaaag cgcatgctcc agactgcctt gggaaaagcg
     5401 cctcccctac ccggtagaat tc
//
