LOCUS       sequence-226719-        6754 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of ARL13b.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6754)
  TITLE     Kahn expression plasmids
REFERENCE   2  (bases 1 to 6754)
  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"
COMMENT     Sequence Label: LN92
FEATURES             Location/Qualifiers
     source          1..6754
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        153..532
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        533..736
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     687..707
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     781..800
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        781..799
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2125..2151
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             2176..2205
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             2221..2238
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2261..2278)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2267..2491
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2537..2965
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2624..2643)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2834..2855
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2974..2994)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2979..3308
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3159..3294
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3221..3240
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3375..4169
                     /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(3429..3448)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4039..4058
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4345..4466
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4382..4401)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4436..4455
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4515..4531)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4515..4531)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4528..4550)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4539..4555
                     /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(4563..4569,4570..4587,4588..4593))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4608..4629
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4746..4763)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4917..5505)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4997..5016)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5676..6467,6468..6536))
                     /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     6299..6318
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6537..6641)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6716..6735)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
       61 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agggttaggc gttttgcgct
      121 gcttcgcgat gtacgggcca gatatacgcg ttgacattga ttattgacta gttattaata
      181 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
      241 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
      301 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
      361 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
      421 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
      481 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtgatgcg
      541 gttttggcag tacatcaatg ggcgtggata gcggtttgac tcacggggat ttccaagtct
      601 ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa aatcaacggg actttccaaa
      661 atgtcgtaac aactccgccc cattgacgca aatgggcggt aggcgtgtac ggtgggaggt
      721 ctatataagc agagctctct ggctaactag agaacccact gcttactggc ttatcgaaat
      781 taatacgact cactataggg agacccaagc tggctagtta agcttggtac cgagctcgga
      841 tccatgttca gtctgatggc caactgctgc aacttgttca agcggtggcg ggagcctgtc
      901 agaaaggtga cacttgtgat ggtcggcctt gataatgctg gtaaaacagc cacagcaaag
      961 ggaatccaag gagagcatcc tgaagacgta gctcccactg ttggcttttc taaaattgat
     1021 ctgagacaag gaaagttcca agttaccatc tttgacttag gaggtggaaa aagaattcgg
     1081 ggaatttgga agaattatta tgctgaatcc tatggggtaa tatttgttgt ggattccagt
     1141 gatgaggaga gaatggaaga aacaaaggag acaatgtccg aagtgctaag acacccgagg
     1201 atatcaggaa agcctatatt ggtgctggcc aacaagcaag acaaggaggg ggctttaggc
     1261 gaagcggatg tgattgagtg tctctcgctg gagaagctgg tcaacgagca caagtgcctg
     1321 tgtcagatcg aaccttgttc agcagtcttg ggatatggaa agaaaattga caagtccatt
     1381 aaaaaggggc tttattggct actgcatatc attgcaaagg actttgatgc cttaagtgaa
     1441 cgcatccaga aagacacaac tgaacagcga gctcttgagg aacaagagaa acgtgagagg
     1501 gctgaacgag tccggaagtt aagggaagaa agagaacgag agcagacgga acttgatggg
     1561 accagtggtc tggctgagat tgactcagga ccagttcttg cgaatccttt ccaacccatt
     1621 gcagctgtaa tcattgagaa tgaaaagaag caggaaaaag agaaaaagaa gcagaccgtg
     1681 gagaaggaca gtgatgttgg tctcctagaa cataaagtgg agccagagca agcagcgcct
     1741 cagagtgagg cagattgctg cctccaaaac cctgacgagc gagtggtgga cagttacagg
     1801 gaggcactgt cgcagcagct ggacagtgag gatgagcagg accagcgagg gtcagagtcc
     1861 ggggaaaaca gtaagaagaa aaccaagaaa ctaagaatga aaaggagtca tcgggtagaa
     1921 ccagtcaata cagacgagtc tactccaaag agtcccacgc ctccccaacc tccccctcct
     1981 gttggctggg gaacccccaa agtcactaga cttccaaaac ttgagcctct tggtgaaaca
     2041 cgtcacaatg atttctatgg gaagccgctg cctcccctgg ctgtgcgaca gagacctaac
     2101 ggtgatgctc aggacacgat ctcataccca tacgatgttc cagattacgc ttaactcgag
     2161 tctagagggc ccttcgaaca aaaactcatc tcagaagagg atctgaatat gcataccggt
     2221 catcatcacc atcaccattg agtttaaacc cgctgatcag cctcgactgt gccttctagt
     2281 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     2341 cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat
     2401 tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc
     2461 aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc
     2521 tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     2581 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     2641 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     2701 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     2761 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     2821 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     2881 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     2941 atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa
     3001 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     3061 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     3121 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     3181 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     3241 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     3301 tttgcaaaaa gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat
     3361 gaggatcgtt tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg
     3421 tggagaggct attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg
     3481 tgttccggct gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg
     3541 ccctgaatga actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc
     3601 cttgcgcagc tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg
     3661 aagtgccggg gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca
     3721 tggctgatgc aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc
     3781 aagcgaaaca tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg
     3841 atgatctgga cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg
     3901 cgcgcatgcc cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata
     3961 tcatggtgga aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg
     4021 accgctatca ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat
     4081 gggctgaccg cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct
     4141 tctatcgcct tcttgacgag ttcttctgag cgggactctg gggttcgcga aatgaccgac
     4201 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     4261 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     4321 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     4381 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     4441 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     4501 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     4561 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     4621 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     4681 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     4741 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     4801 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     4861 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     4921 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     4981 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     5041 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     5101 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     5161 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     5221 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     5281 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     5341 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     5401 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     5461 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     5521 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     5581 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     5641 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     5701 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     5761 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     5821 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     5881 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     5941 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     6001 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     6061 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     6121 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     6181 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     6241 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     6301 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     6361 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     6421 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     6481 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     6541 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     6601 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     6661 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     6721 tctgctctga tgccgcatag ttaagccagt atct
//
