LOCUS       sequence-424507-        7464 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 7464)
  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 7464)
  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: pDEST47-Arl13b12
FEATURES             Location/Qualifiers
     source          1..7464
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        160..539
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        540..743
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     694..714
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     788..807
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        788..806
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    839..863
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     RBS             875..895
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    complement(2202..2226)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             join(2237..2239,2240..2242,2243..2956)
                     /codon_start=1
                     /product="Cycle 3 GFP (Crameri et al., 1996)"
                     /label=Cycle 3 GFP
                     /note="mammalian codon-optimized"
                     /translation="MASKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFSYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITHGMDELYK"
     primer_bind     complement(2270..2298)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2893..2913
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2978..2995)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2984..3208
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3254..3682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3341..3360)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3551..3572
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3691..3711)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3696..4025
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3876..4011
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3938..3957
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4092..4886
                     /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(4146..4165)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4756..4775
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5062..5183
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5099..5118)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5153..5172
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5232..5248)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5232..5248)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5245..5267)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5256..5272
                     /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(5280..5286,5287..5304,5305..5310))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5325..5346
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5463..5480)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5634..6222)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5714..5733)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6393..7184,7185..7253))
                     /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     7016..7035
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7254..7358)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7433..7452)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 agtatctgct ccctgcttgt gtgttggagg tcgctgagta gtgcgcgagc aaaatttaag
       61 ctacaacaag gcaaggcttg accgacaatt gcatgaagaa tctgcttagg gttaggcgtt
      121 ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta ttgactagtt
      181 attaatagta atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta
      241 cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt
      301 caataatgac gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg
      361 tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta
      421 cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga
      481 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg
      541 tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc
      601 caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact
      661 ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt
      721 gggaggtcta tataagcaga gctctctggc taactagaga acccactgct tactggctta
      781 tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagttaagc ttgatcaaac
      841 aagtttgtac aaaaaagcag gctccgcggc cgccttgttt aactttaaga aggagccctt
      901 caccatgttc agtctgatgg ccaactgctg caacttgttc aagcggtggc gggagcctgt
      961 cagaaaggtg acacttgtga tggtcggcct tgataatgct ggtaaaacag ccacagcaaa
     1021 gggaatccaa ggagagcatc ctgaagacgt agctcccact gttggctttt ctaaaattga
     1081 tctgagacaa ggaaagttcc aagttaccat ctttgactta ggaggtggaa aaagaattcg
     1141 gggaatttgg aagaattatt atgctgaatc ctatggggta atatttgttg tggattccag
     1201 tgatgaggag agaatggaag aaacaaagga gacaatgtcc gaagtgctaa gacacccgag
     1261 gatatcagga aagcctatat tggtgctggc caacaagcaa gacaaggagg gggctttagg
     1321 cgaagcggat gtgattgagt gtctctcgct ggagaagctg gtcaacgagc acaagtgcct
     1381 gtgtcagatc gaaccttgtt cagcagtctt gggatatgga aagaaaattg acaagtccat
     1441 taaaaagggg ctttattggc tactgcatat cattgcaaag gactttgatg ccttaagtga
     1501 acgcatccag aaagacacaa ctgaacagcg agctcttgag gaacaagaga aacgtgagag
     1561 ggctgaacga gtccggaagt taagggaaga aagagaacga gagcagacgg aacttgatgg
     1621 gaccagtggt ctggctgaga ttgactcagg accagttctt gcgaatcctt tccaacccat
     1681 tgcagctgta atcattgaga atgaaaagaa gcaggaaaaa gagaaaaaga agcagaccgt
     1741 ggagaaggac agtgatgttg gtctcctaga acataaagtg gagccagagc aagcagcgcc
     1801 tcagagtgag gcagattgct gcctccaaaa ccctgacgag cgagtggtgg acagttacag
     1861 ggaggcactg tcgcagcagc tggacagtga ggatgagcag gaccagcgag ggtcagagtc
     1921 cggggaaaac agtaagaaga aaaccaagaa actaagaatg aaaaggagtc atcgggtaga
     1981 accagtcaat acagacgagt ctactccaaa gagtcccacg cctccccaac ctccccctcc
     2041 tgttggctgg ggaaccccca aagtcactag acttccaaaa cttgagcctc ttggtgaaac
     2101 acgtcacaat gatttctatg ggaagccgct gcctcccctg gctgtgcgac agagacctaa
     2161 cggtgatgct caggacacga tctcaaaggg tgggcgcgcc gacccagctt tcttgtacaa
     2221 agtggttcga tctagaatgg ctagcaaagg agaagaactt ttcactggag ttgtcccaat
     2281 tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg gagagggtga
     2341 aggtgatgct acatacggaa agcttaccct taaatttatt tgcactactg gaaaactacc
     2401 tgttccatgg ccaacacttg tcactacttt ctcttatggt gttcaatgct tttcccgtta
     2461 tccggatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     2521 ggaacgcact atatctttca aagatgacgg gaactacaag acgcgtgctg aagtcaagtt
     2581 tgaaggtgat acccttgtta atcgtatcga gttaaaaggt attgatttta aagaagatgg
     2641 aaacattctc ggacacaaac tcgagtacaa ctataactca cacaatgtat acatcacggc
     2701 agacaaacaa aagaatggaa tcaaagctaa cttcaaaatt cgccacaaca ttgaagatgg
     2761 atccgttcaa ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct
     2821 tttaccagac aaccattacc tgtcgacaca atctgccctt tcgaaagatc ccaacgaaaa
     2881 gcgtgaccac atggtccttc ttgagtttgt aactgctgct gggattacac atggcatgga
     2941 tgagctctac aaataatgaa ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc
     3001 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     3061 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     3121 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     3181 catgctgggg atgcggtggg ctctatggct tctgaggcgg aaagaaccag ctggggctct
     3241 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3301 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3361 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta
     3421 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     3481 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     3541 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     3601 tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt
     3661 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     3721 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3781 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     3841 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     3901 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     3961 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4021 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4081 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4141 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4201 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4261 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4321 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4381 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4441 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     4501 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     4561 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     4621 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     4681 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     4741 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     4801 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     4861 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     4921 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     4981 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5041 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5101 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5161 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5221 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5281 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5341 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5401 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     5461 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     5521 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     5581 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     5641 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     5701 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     5761 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     5821 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     5881 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     5941 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6001 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6061 ggctacacta gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6121 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6181 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6241 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6301 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6361 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6421 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     6481 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     6541 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     6601 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     6661 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     6721 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     6781 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     6841 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     6901 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     6961 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7021 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7081 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7141 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7201 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7261 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7321 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7381 cctgacgtcg acggatcggg agatctcccg atcccctatg gtgcactctc agtacaatct
     7441 gctctgatgc cgcatagtta agcc
//
