LOCUS       sequence-348618-        6829 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6829)
  AUTHORS   Larkins CE, Aviles GD, East MP, Kahn RA, Caspary T
  TITLE     Arl13b regulates ciliogenesis and the dynamic localization of Shh 
            signaling proteins.
  JOURNAL   Mol Biol Cell. 2011 Dec;22(23):4694-703. Epub 2011 Oct 5.
  PUBMED    21976698
REFERENCE   2  (bases 1 to 6829)
  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: "Mol Biol 
            Cell."; date: "2011-12"; volume: "22(23)"; pages: "4694-703. Epub 
            2011 Oct 5"
COMMENT     Sequence Label: pDEST40-Arl13b89
FEATURES             Location/Qualifiers
     source          1..6829
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    677..701
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             713..733
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2043..2067)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             2093..2134
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             2144..2161
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2184..2201)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2190..2414
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2460..2888
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2547..2566)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2757..2778
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2897..2917)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2902..3231
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3082..3217
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3144..3163
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3298..4092
                     /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(3352..3371)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3962..3981
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4268..4389
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4305..4324)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4359..4378
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4438..4454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4438..4454)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4451..4473)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4462..4478
                     /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(4486..4492,4493..4510,4511..4516))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4531..4552
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4669..4686)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4840..5428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4920..4939)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5599..6390,6391..6459))
                     /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     6222..6241
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6460..6564)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6639..6658)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagttaag ctatcaacaa gtttgtacaa aaaagcaggc tccgcggccg ccttgtttaa
      721 ctttaagaag gagcccttca ccatgttcag tctgatggcc aactgctgca acttgttcaa
      781 gcggtggcgg gagcctgtca gaaaggtgac acttgtgatg gtcggccttg ataatgctgg
      841 taaaacagcc acagcaaagg gaatccaagg agagcatcct gaagacgtag ctcccactgt
      901 tggcttttct aaaattgatc tgagacaagg aaagttccaa gttaccatct ttgacttagg
      961 aggtggaaaa agaattcggg gaatttggaa gaattattat gctgaatcct atggggtaat
     1021 atttgttgtg gattccagtg atgaggagag aatggaagaa acaaaggaga caatgtccga
     1081 agtgctaaga cacccgagga tatcaggaaa gcctatattg gtgctggcca acaagcaaga
     1141 caaggagggg gctttaggcg aagcggatgt gattgagtgt ctctcgctgg agaagctggt
     1201 caacgagcac aagtgcctgt gtcagatcga accttgttca gcagtcttgg gatatggaaa
     1261 gaaaattgac aagtccatta aaaaggggct ttattggcta ctgcatatca ttgcaaagga
     1321 ctttgatgcc ttaagtgaac gcatccagaa agacacaact gaacagcgag ctcttgagga
     1381 acaagagaaa cgtgagaggg ctgaacgagt ccggaagtta agggaagaaa gagaacgaga
     1441 gcagacggaa cttgatggga ccagtggtct ggctgagatt gactcaggac cagttcttgc
     1501 gaatcctttc caacccattg cagctgtaat cattgagaat gaaaagaagc aggaaaaaga
     1561 gaaaaagaag cagaccgtgg agaaggacag tgatgttggt ctcctagaac ataaagtgga
     1621 gccagagcaa gcagcgcctc agagtgaggc agattgctgc ctccaaaacc ctgacgagcg
     1681 agtggtggac agttacaggg aggcactgtc gcagcagctg gacagtgagg atgagcagga
     1741 ccagcgaggg tcagagtccg gggaaaacag taagaagaaa accaagaaac taagaatgaa
     1801 aaggagtcat cgggtagaac cagtcaatac agacgagtct actccaaaga gtcccacgcc
     1861 tccccaacct ccccctcctg ttggctgggg aacccccaaa gtcactagac ttccaaaact
     1921 tgagcctctt ggtgaaacac gtcacaatga tttctatggg aagccgctgc ctcccctggc
     1981 tgtgcgacag agacctaacg gtgatgctca ggacacgatc tcataaaagg gtgggcgcgc
     2041 cgacccagct ttcttgtaca aagtggttga tctagagggc ccgcggttcg aaggtaagcc
     2101 tatccctaac cctctcctcg gtctcgattc tacgcgtacc ggtcatcatc accatcacca
     2161 ttgagtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2221 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2281 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2341 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2401 ggtgggctct atggcttctg aggcggaaag aaccagctgg ggctctaggg ggtatcccca
     2461 cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg gttacgcgca gcgtgaccgc
     2521 tacacttgcc agcgccctag cgcccgctcc tttcgctttc ttcccttcct ttctcgccac
     2581 gttcgccggc tttccccgtc aagctctaaa tcgggggctc cctttagggt tccgatttag
     2641 tgctttacgg cacctcgacc ccaaaaaact tgattagggt gatggttcac gtagtgggcc
     2701 atcgccctga tagacggttt ttcgcccttt gacgttggag tccacgttct ttaatagtgg
     2761 actcttgttc caaactggaa caacactcaa ccctatctcg gtctattctt ttgatttata
     2821 agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac aaaaatttaa
     2881 cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca
     2941 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc
     3001 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
     3061 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
     3121 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc tgcctctgag
     3181 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctcccg
     3241 ggagcttgta tatccatttt cggatctgat caagagacag gatgaggatc gtttcgcatg
     3301 attgaacaag atggattgca cgcaggttct ccggccgctt gggtggagag gctattcggc
     3361 tatgactggg cacaacagac aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg
     3421 caggggcgcc cggttctttt tgtcaagacc gacctgtccg gtgccctgaa tgaactgcag
     3481 gacgaggcag cgcggctatc gtggctggcc acgacgggcg ttccttgcgc agctgtgctc
     3541 gacgttgtca ctgaagcggg aagggactgg ctgctattgg gcgaagtgcc ggggcaggat
     3601 ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca tcatggctga tgcaatgcgg
     3661 cggctgcata cgcttgatcc ggctacctgc ccattcgacc accaagcgaa acatcgcatc
     3721 gagcgagcac gtactcggat ggaagccggt cttgtcgatc aggatgatct ggacgaagag
     3781 catcaggggc tcgcgccagc cgaactgttc gccaggctca aggcgcgcat gcccgacggc
     3841 gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga atatcatggt ggaaaatggc
     3901 cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata
     3961 gcgttggcta cccgtgatat tgctgaagag cttggcggcg aatgggctga ccgcttcctc
     4021 gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg ccttctatcg ccttcttgac
     4081 gagttcttct gagcgggact ctggggttcg cgaaatgacc gaccaagcga cgcccaacct
     4141 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4201 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4261 ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     4321 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     4381 tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt aatcatggtc
     4441 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     4501 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     4561 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     4621 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     4681 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     4741 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     4801 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     4861 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     4921 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     4981 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     5041 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     5101 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     5161 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     5221 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     5281 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     5341 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     5401 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     5461 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     5521 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     5581 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     5641 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     5701 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     5761 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     5821 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     5881 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     5941 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     6001 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     6061 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     6121 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     6181 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     6241 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     6301 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     6361 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     6421 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     6481 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     6541 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtcgacgg
     6601 atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc tgatgccgca
     6661 tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc
     6721 aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg
     6781 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
