LOCUS       sequence-67410-e        7504 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AU1 epitope-tagged TTC30B in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7504)
  AUTHORS   Howard PW, Jue SF, Maurer RA
  TITLE     Interaction of mouse TTC30/DYF-1 with multiple intraflagellar 
            transport complex B proteins and KIF17.
  JOURNAL   Exp Cell Res. 2013 Jun 25. pii: S0014-4827(13)00271-1. doi: 
            10.1016/j.yexcr.2013.06.010.
  PUBMED    23810713
REFERENCE   2  (bases 1 to 7504)
  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: "Exp Cell 
            Res. 2013 Jun 25. pii: S0014-4827(13)00271-1. doi: 
            10.1016/j.yexcr.2013.06.010."
COMMENT     Sequence Label: AU1-TTC30B-pcDNA3
FEATURES             Location/Qualifiers
     source          1..7504
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     911..927
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     912..928
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(3056..3074)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3057..3074)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3094..3111)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3100..3324
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3370..3798
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3457..3476)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3667..3688
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3807..3827)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3812..4142
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3993..4128
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4055..4074
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4209..5003
                     /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(4263..4282)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4873..4892
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5177..5298
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5214..5233)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5268..5287
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5347..5363)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5347..5363)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5360..5382)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5371..5387
                     /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(5395..5401,5402..5419,5420..5425))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5440..5461
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5578..5595)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5749..6337)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5829..5848)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6508..7299,7300..7368))
                     /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     7131..7150
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7369..7473)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggac tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc
      901 gggccccccc tcgaggtcga cggtatcgat aagcttgcca acatggcaga cacataccgc
      961 tacatctcca tggcagggct gagcagctca cagatccccg acggtgagtt cacagcagtg
     1021 gtgtacaggc tcatccgcga ctcccgctac tcagaggcgg tgcagctgct gagcgcagag
     1081 ctgcagagga gctcccgcag ccgcgcaggg ctgtcgctgc tggcctactg ctactaccgc
     1141 ctgcaggagt tcgagctcgc tgcagagtgc tatgagcagc tgagccagat gcacccggaa
     1201 ctcgagcagt accgcctgta ccaggcccag gcactgtaca aggcctgcct gtatccagag
     1261 gccaccaggg tcgccttcct cctggacaac cccgcctatc agactcgagt ccttcgtctc
     1321 caggctgcta tcaagtacag tgaaggtgac ctcccaggag ccaggagcct ggtggagcag
     1381 ctgctgagtg gggaaggtgt agaggacagt ggaggggaga gtgattatga tggtcagatc
     1441 aacctaggtt gtctgctcta caaagaggga cactatgaag ctgcctgttc caagttctta
     1501 gcggccctgc aggcttctgg ctaccagcct gacctttcct acaacttggc tttggcctat
     1561 tacagcagtc ggcagtacgc gcctgctctg aagcatatcg ctgacatcat tgagcgtggc
     1621 atccgtcaac acccagaact cggtgtgggc atgaccacgg agggcattga tgttcgcagt
     1681 gttggcaaca ctatagtcct tcaccagact gctctggtcg aagccttcaa cctcaaggca
     1741 gccatagagt accagctgag aaactacgag gcagcccagg aagccctcac tgacatgcca
     1801 cccagagcag aggaagagct ggaccccgta accctccaca accaggctct gatgaacatg
     1861 gatgccaagc ccacagaggg ctttgaaaag ctccagtttc tgctccagca gaaccccttc
     1921 cccccagaga cctttggcaa cctgctgctg ctctactgta aatatgagta ttttgacctg
     1981 gcagctgatg tcctggcaga aaatgcccac ttgacttaca agttcctcac accctatctc
     2041 tatgacttct tggatgccat gatcacttgc cagacagctc ctgaagaggc tttcattaaa
     2101 cttgatgggt tggctggcat gctgactgag cagctccgga aactcaccat acaagtacag
     2161 gattcaaggc atagtagaga tgatgaatct gccaaaaagg ctgtgaatga atatgatgaa
     2221 actcttgaga agtatattcc tgtactgatg gcccaggcaa aaatctactg gaactttgag
     2281 aattacccaa tggtggagaa gatcttccgc aaatctgtgg aattctgtaa tgaccatgat
     2341 gtgtggaagc tgaatgtggc ccatgtcctc ttcatgcagg agaacaagta caaagaggcc
     2401 attggtttct atgagcccat tgtcaagaag aattatgata acatcctgag tgtcagtgct
     2461 attgtgttag ccaacctctg tgtttcctat atcatgacaa gtcaaaatga ggaagctgag
     2521 gagctcatga ggaagattga gaaggaggag gaacaactct cttatggtga cccagacaag
     2581 aaaatctacc acctttgcat tgtgaatttg gtgataggaa ctctgtattg tgccaaagga
     2641 aactatgact ttggcatctc tcgggttatc aaaagcctgg agccttacca taaaaagcta
     2701 ggcaccgata cgtggtatta tgccaagaga tgtttcctgt ccttactaga gaacatgtca
     2761 aagcacacaa ttatgctaag ggacagtgtc attcaagaat gtgtgcagtt tctagagcac
     2821 tgtgaacttt atggtagaaa cattccagct gttatagaac agcccttgga ggaagaaaga
     2881 atgcacactg ggaagaatac agtcacatat gaatcaagaa agttgagggc tctgatttat
     2941 gagataattg gatggaatgt gtaaggatcc actagtaacg gccgccagtg tgctggaatt
     3001 ctgcagatat ccatcacact ggcggccgct cgagcatgca tctagagggc cctattctat
     3061 agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct agttgccagc
     3121 catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg
     3181 tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc
     3241 tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg
     3301 ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg
     3361 ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca
     3421 gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct
     3481 ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc cctttagggt
     3541 tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac
     3601 gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct
     3661 ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt
     3721 ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag ctgatttaac
     3781 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3841 aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3901 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3961 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     4021 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     4081 ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     4141 aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca ggatgaggat
     4201 cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct tgggtggaga
     4261 ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc gccgtgttcc
     4321 ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc ggtgccctga
     4381 atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc gttccttgcg
     4441 cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg ggcgaagtgc
     4501 cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc atcatggctg
     4561 atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac caccaagcga
     4621 aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat caggatgatc
     4681 tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc aaggcgcgca
     4741 tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg aatatcatgg
     4801 tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg gcggaccgct
     4861 atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc gaatgggctg
     4921 accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc gccttctatc
     4981 gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg accaagcgac
     5041 gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt
     5101 cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga
     5161 gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     5221 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     5281 actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta
     5341 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     5401 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     5461 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     5521 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc
     5581 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5641 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5701 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5761 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5821 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5881 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5941 tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     6001 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     6061 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     6121 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6181 cactagaagg acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6241 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6301 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6361 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6421 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6481 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6541 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6601 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6661 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6721 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6781 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6841 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6901 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6961 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     7021 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     7081 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     7141 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7201 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7261 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7321 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7381 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7441 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7501 cgtc
//
