LOCUS       sequence-401481-        9577 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Doxycycline inducible expression of CDK5RAP2 cDNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9577)
  AUTHORS   Nadler MJS, Chang W, Ozkaynak E, Huo Y, Nong Y, Boillot M, Johnson 
            M, Moreno A, Matthew P Anderson
  TITLE     Hominoid SVA-lncRNA AK057321 targets human-specific SVA 
            retrotransposons in SCN8A and CDK5RAP2 to initiate neuronal 
            maturation.
  JOURNAL   Commun Biol. 2023 Mar 30;6(1):347. doi: 10.1038/s42003-023-04683-8.
  PUBMED    36997626
REFERENCE   2  (bases 1 to 9577)
  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: "Commun 
            Biol."; date: "2023-03-30"; volume: "6(1):347. doi"; pages: " 
            10.1038/s42003-023-04683-8"
COMMENT     Sequence Label: TetO-CDK5RAP2
FEATURES             Location/Qualifiers
     source          1..9577
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    17..35
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    59..77
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    101..119
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    143..161
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     promoter        194..232
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     CDS             341..370
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     RBS             1115..1126
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     polyA_signal    6236..6357
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6273..6292)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6327..6346
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6364..6819)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6501..6522)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6713..6732
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        6846..6950
                     /gene="bla"
                     /label=AmpR promoter
     promoter        6952..7309
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        6952..7143
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(6975..6995)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      7160..7295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             7344..8138
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             7344..8138
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(7398..7417)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     8008..8027
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(8326..8345)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    8370..8417
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8746..9334
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9235..9254
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     misc_feature    9420..9537
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
ORIGIN
        1 atctcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
       61 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
      121 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagctcg
      181 gtacccgggt cgaggtaggc gtgtacggtg ggaggcctat ataagcagag ctcgtttagt
      241 gaaccgtcaa gcgctaccgg actcagatct cgagctcaag cttgcttaca tttgcttctg
      301 acacaactgt gttcactagc aacctcaaac agacaccatg gagcagaagc tgatctccga
      361 ggaggacctg aacatgaatt cggccggcct gaccatgatg gacttggtgt tggaagagga
      421 cgtcaccgtc cctgggacgc tcagcggctg cagtggcctt gttcccagtg taccagatga
      481 cctggatggc atcaacccca atgctgggtt gggaaatggt ctgctcccaa atgtgtcaga
      541 agaaacagtg tctcccacca gagcacggaa catgaaggac tttgaaaatc aaatcactga
      601 attgaagaaa gaaaacttta acctaaagct ccgcatctat ttccttgagg aaagaatgca
      661 acaggaattt catggcccca ctgaacatat ctacaaaact aacattgagc tcaaggtgga
      721 agtagaaagt ctgaagcggg aactccagga gagagagcag ctgctcatca aagcctccaa
      781 agcagttgag agcttagctg aagcaggtgg ctctgaaatc cagcgggtga aagaagatgc
      841 tcgaaagaag gtgcagcagg tggaagatct cctaactaaa agaatactcc ttttggaaaa
      901 ggatgtgaca gccgcccagg cagaactgga aaaggccttt gcagggacag agacggagaa
      961 ggctcttcgg ttgcgtttgg aaagcaagct ttcagagatg aagaagatgc acgaggggga
     1021 cttggcgatg gctctggtcc tggatgagaa agacagactg attgaggagt tgaagctgtc
     1081 tttgaagagc aaagaagctt taattcagtg ccttaaagag gagaaatctc agatggcatg
     1141 tcctgatgag aatgtgtcat ctggagagct ccgaggactt tgtgctgctc caagggaaga
     1201 aaaggagaga gaaactgagg ctgcacaaat ggagcatcag aaggagagaa acagctttca
     1261 agagaggatc caggcacttg aagaggacct gagagagaag gaaagagaaa ttgctacaga
     1321 gaagaaaaat agtctaaaga gggataaagc cattcagggt ttaaccatgg cattaaaatc
     1381 aaaggaaaaa aaggttgaag aacttaactc tgaaattgaa aagctcagtg ctgcctttgc
     1441 taaagccaga gaggccctac agaaagcaca gacccaggaa tttcaggggt ctgaagacta
     1501 tgagactgct ctatcaggaa aggaagccct ttcggctgcg ctgcgctcac aaaacctcac
     1561 caagagtaca gagaaccaca gactgcgtag aagcattaag aagatcaccc aggagctgag
     1621 tgacttgcag caggagaggg agagactgga gaaggacctg gaggaagccc atcgagagaa
     1681 gagcaaagga gactgcacca tccgtgatct tagaaatgaa gttgaaaaat tacgcaatga
     1741 agtgaatgaa agagagaaag caatggaaaa tcgttacaag agtcttctga gtgaaagcaa
     1801 taaaaaattg cacaatcaag agcaagtgat caaacatcta acagaaagta ccaatcagaa
     1861 ggacgtgttg cttcagaaat tcaatgaaaa agatttggaa gtaatacagc agaactgcta
     1921 tttaatggct gcagaggatc ttgagctcag gagtgaaggc ttaataacag aaaagtgctc
     1981 ttctcaacag ccaccaggca gcaaaaccat cttctctaag gaaaagaaac aatcatcaga
     2041 ctatgaagag ctgattcagg tcttaaagaa agagcaggac atctataccc atctggtcaa
     2101 atctctgcag gaatcagaca gtatcaacaa cctgcaggct gagttaaaca agatttttgc
     2161 cctgcggaag caactggagc aggatgtgct ttcatatcag aatttgcgga agaccctcga
     2221 ggagcagatc agcgaaattc ggaggcggga agaagaatca ttttcacttt atagtgatca
     2281 aacattttat ctaagtattt gccttgaaga aaacaatcgg tttcaagtgg aacatttttc
     2341 tcaagaagaa cttaagaaaa aggtcagtga ccttatacag ctagtgaagg agctgtatac
     2401 agacaaccag cacctgaaga aaaccatttt tgatctctcc tgcatgggtt tccagggaaa
     2461 tgggtttcca gatagacttg cgtctacaga acaaacagag cttctggcta gcaaggagga
     2521 cgaggacacg atcaaaattg gggaggatga cgagattaat ttcctgagtg accagcattt
     2581 gcagcagagt aatgagatta tgaaagacct ttccaaagga ggctgcaaaa atggatactt
     2641 aaggcacacg gagtctaaga tttcagattg tgatggggcc cacgcacctg gctgcctaga
     2701 agaaggtgca ttcataaacc tgcttgcccc tttgttcaat gagaaggcca cattattact
     2761 ggaatccagg ccagaccttc tgaaagtggt acgggaactg cttctgggac aactattctt
     2821 gacagagcag gaagtttctg gagaacacct tgatggtaaa actgagaaga cacctaagca
     2881 aaaaggtgaa cttgtacatt ttgtccaaac caactcattt tccaagccac atgatgaact
     2941 gaagttgtct tgtgaggccc agctagtaaa ggcaggcgaa gtgcccaagg taggactgaa
     3001 agatgcctca gtgcagactg tggccacgga gggcgacctg ctgagattca agcatgaagc
     3061 aacaagagag gcttgggaag agaaaccgat caacactgca ctcagcgcag agcatcggcc
     3121 agagaacctg cacggggtgc ctgggtggca ggctgccctc ctttccctcc ctggtattac
     3181 caacagagag gctaagaagt cccgcttgcc aatcctaata aaaccatccc ggtcattagg
     3241 aaatatgtat cgtctccctg ccacccagga ggtggtgacg cagctgcaga gccagatctt
     3301 ggagctgcag ggggagctga aggagtttaa aacttgtaat aagcaacttc accaaaagtt
     3361 aattctggct gaagcagtga tggaggggag gccaacgccc gacaaaacgt tgctgaatgc
     3421 tcagccccct gtgggagcag cctaccagga cagcccagga gagcagaaag gaattaaaac
     3481 cacatcttct gtctggagag acaaggaaat ggacagtgat cagcaaagaa gctacgagat
     3541 tgactctgag atttgcccac ctgatgacct tgccagcttg ccatcatgca aagaaaatcc
     3601 tgaagatgtt ctgagcccaa cttcagtagc tacttacctg agttccaaga gtcagccttc
     3661 tgctaaagtc agtgtgatgg ggactgatca gtcagagagc attaatacct caaatgagac
     3721 agaatactta aaacagaaaa tccatgactt ggaaactgag ctggaaggct accagaattt
     3781 catatttcag cttcaaaagc actcccagtg cagtgaggcc ataattacag ttttgtgtgg
     3841 gacagaaggg gcccaggatg gcttgagcaa gcccaagaat ggttctgatg gggaagaaat
     3901 gaccttttca agtttgcacc aagtgcgata cgtgaaacac gtgaaaatcc tcggtccgct
     3961 ggccccagag atgattgaca gcagggtgct ggagaacctc aaacagcagc tggaggaaca
     4021 ggaatacaag ctgcagaagg agcagaattt gaacatgcaa cttttcagtg agatccataa
     4081 tctgcagaat aagttcagag atctctcacc tcccagatac gattcattag ttcagtccca
     4141 agccagggag ctctcccttc aacggcagca gattaaggat ggccatggca tctgtgtcat
     4201 ctcccgtcaa cacatgaaca ccatgattaa ggcatttgag gagttgctgc aggccagtga
     4261 tgtggattac tgtgtggccg agggtttcca ggaacagctg aatcaatgtg ctgagctgct
     4321 ggagaaattg gaaaagctat ttctcaacgg aaaatcagtt ggagtggaaa tgaacaccca
     4381 gaatgaactg atggagagga ttgaggaaga caacttaacc taccaacatc ttctgcctga
     4441 atctcctgag ccttcagcct ctcatgcgct ctctgattat gaaacatctg aaaagtcctt
     4501 cttctcacga gaccagaagc aagataatga gacagagaag acttcagtta tggtgaacag
     4561 tttttctcaa gacttactaa tggaacacat acaggaaatt cgaactttga gaaagcgttt
     4621 agaagaatct attaaaacaa atgagaagct acggaaacag ttggaacggc aaggatctga
     4681 atttgttcaa ggttctacaa gcatttttgc ttctggttca gagcttcata gttctctaac
     4741 atcagaaatt catttcttga ggaagcagaa ccaggccctc aatgcaatgc tcattaaagg
     4801 atccagagat aaacagaagg agaatgacaa attacgagag tccctctcca ggaagaccgt
     4861 gagcctggag caccttcagc gggagtatgc cagcgtgaag gaagaaaatg aaaggctgca
     4921 gaaagaaggc agcgagaagg agagacacaa ccagcagctg atccaggagg tccgctgcag
     4981 cggccaggag ctgagcaggg tgcaggagga gctgaagttg aggcagcagc tgctctcaca
     5041 gaatgacaag ctattgcagt ctctccgagt ggagctgaag gcgtatgaga agctggatga
     5101 agagcacagg agactgagag aggcgtcggg agaaggctgg aaggggcagg atcctttcag
     5161 ggacctgcac agcctcctga tggagatcca ggctctgcgc ttgcaactag aaaggagcat
     5221 cgaaaccagc agcactctgc agagcaggct caaggaacag ctggcaaggg gggcagagaa
     5281 ggcacaggaa ggagccctca ctctggctgt ccaagccgtg tccatccctg aggtgcccct
     5341 tcagcctgac aaacacgatg gtgacaaata tcccatggaa agtgataatt catttgatct
     5401 gtttgattcc tcccaggcag tgacaccaaa atcagtttca gagactcctc cactctctgg
     5461 gaatgacacg gactccctct cctgcgacag tggcagttcg gcaactagca ctccgtgtgt
     5521 gtcccgcctg gtcactggcc accacctgtg ggccagcaag aatggccgcc atgtcctggg
     5581 cctgattgag gactatgagg ccctgctcaa acagatcagc cagggacaga ggctccttgc
     5641 tgaaatggac attcaaaccc aagaggctcc cagctccaca agtcaagagc tgggaacaaa
     5701 gggtccacac ccagcaccac tgagcaagtt tgtgagcagt gtgagcacgg ccaagctgac
     5761 cctggaagag gcctacaggc ggctgaagct tctctggaga gtctcactcc ccgaggatgg
     5821 ccagtgcccc cttcactgtg agcagattgg agaaatgaag gcagaggtca ccaaactaca
     5881 taaaaaattg tttgaacaag aaaagaagtt gcaaaacacc atgaagcttt tgcagctgag
     5941 caagcgccag gaaaaagtca tctttgatca attggtcgta acccacaaaa tccttcggaa
     6001 ggccagagga aacctggagc ttaggcctgg gggagcccat ccaggaacat gcagtcccag
     6061 cagaccaggc tccgcggccg ctgcagtcga cggtaccgcg ggcccgggat ccaccggatc
     6121 tagataactg atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     6181 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     6241 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     6301 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaacg
     6361 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     6421 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     6481 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     6541 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     6601 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     6661 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     6721 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     6781 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     6841 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     6901 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     6961 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     7021 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     7081 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     7141 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     7201 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     7261 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     7321 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     7381 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     7441 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     7501 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     7561 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     7621 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     7681 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     7741 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     7801 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     7861 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     7921 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     7981 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     8041 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     8101 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     8161 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     8221 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     8281 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     8341 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     8401 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     8461 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     8521 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     8581 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     8641 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     8701 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     8761 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     8821 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     8881 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     8941 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     9001 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     9061 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     9121 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     9181 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     9241 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     9301 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     9361 gttcctggcc ttttgctggc cttttgctca catgtacaaa tggcagtatt catccacaat
     9421 tttaaaagaa aaggggggat tggggggtac agtgcagggg aaagaatagt agacataata
     9481 gcaacagaca tacaaactaa agaattacaa aaacaaatta caaaaattca aaattttcgg
     9541 gtttattaca gggacagcag agatccagtt tggttag
//
