LOCUS       sequence-178304-        7181 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFLAG-CMV2 with cDNA encoding HsNPHP4 aa residues 1-839.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7181)
  AUTHORS   Schou KB, Mogensen JB, Morthorst SK, Nielsen BS, Aleliunaite A, 
            Serra-Marques A, Furstenberg N, Saunier S, Bizet AA, Veland IR, 
            Akhmanova A, Christensen ST, Pedersen LB
  TITLE     KIF13B establishes a CAV1-enriched microdomain at the ciliary 
            transition zone to promote Sonic hedgehog signalling.
  JOURNAL   Nat Commun. 2017 Jan 30;8:14177. doi: 10.1038/ncomms14177.
  PUBMED    28134340
REFERENCE   2  (bases 1 to 7181)
  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: "Nat 
            Commun."; date: "2017-01-30"; volume: "8:14177. doi"; pages: " 
            10.1038/ncomms14177"
COMMENT     Sequence Label: LP453
FEATURES             Location/Qualifiers
     source          1..7181
                     /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"
     CDS             618..641
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    3205..3827
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3404..3423)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3851..3871)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3856..4185
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4036..4171
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4223..4242)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4224..4242)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4256..4272)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4256..4272)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4269..4291)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4280..4296
                     /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(4304..4310,4311..4328,4329..4334))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4349..4370
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4487..4504)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4658..5246)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4738..4757)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5417..6208,6209..6277))
                     /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     6040..6059
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6278..6382)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6409..6864
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6496..6515)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6706..6727
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6990..7012
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7004..7021
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7005..7021
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
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 ccgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttacggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta caccaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaataa ccccgccccg ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agaattgatc taccatggac tacaaagacg atgacgacaa gcttgcggcc gcaatgaacg
      661 actggcacag gattttcacc caaaacgtgc ttgtccctcc ccacccacag agagcgcgcc
      721 agccttggaa ggaatccacg gcattccagt gtgtcctcaa gtggctggac ggaccggtaa
      781 ttaggcaggg cgtgctggag gtactgtcag aggttgaatg ccatctgcga gtgtctttct
      841 ttgatgtcac ctaccggcac ttctttggga ggacgtggaa aaccacagtg aagccgacga
      901 agagaccgcc gtccaggatc gtctttaatg agcccttgta ttttcacaca tccctaaacc
      961 accctcatat cgtggctgtg gtggaagtgg tcgctgaggg caagaaacgg gatgggagcc
     1021 tccagacatt gtcctgtggg tttggaattc ttcggatctt cagcaaccag ccggactctc
     1081 ctatctctgc ttcccaggac aaaaggttgc ggctgtacca tggcaccccc agagccctcc
     1141 tgcacccgct tctccaggac cccgcagagc aaaacagaca catgaccctc attgagaact
     1201 gcagcctgca gtacacgctg aagccacacc cggccctgga gcctgcgttc caccttcttc
     1261 ctgagaacct tctggtgtct ggtctgcagc agatacctgg cctgcttcca gctcatggag
     1321 aatccggcga cgctctccga aagcctcgcc tccagaagcc catcacgggg cacttggatg
     1381 acttattctt caccctgtac ccctccctgg agaagtttga ggaagagctg ctggagctcc
     1441 acgtccagga ccacttccag gagggatgtg gcccactgga cggtggtgcc ctggagatcc
     1501 tggagcggcg cctgcgtgtg ggcgtgcaca atggtctggg cttcgtgcag aggccgcagg
     1561 tcgttgtact ggtgcctgag atggatgtgg ccttgacgcg ctcagctagc ttcagcagga
     1621 aagtggtctc ctcttccaag accagctccg ggagccaagc tctggttttg agaagccgcc
     1681 tccgcctccc agagatggtc ggccaccctg catttgcggt catcttccag ctggagtacg
     1741 tgttcagcag ccctgcagga gtggacggca atgcagcttc ggtcacctct ctgtccaacc
     1801 tggcatgcat gcacatggtc cgctgggctg tttggaaccc cttgctggaa gctgattctg
     1861 gaagggtgac cctgcctctg cagggtggaa tccagcccaa cccctcgcac tgtctggtct
     1921 acaaggtacc ctcagccagc atgagctctg aagaggtgaa gcaggtggag tcgggtacac
     1981 tccggttcca gttctcgctg ggctcagaag aacacctgga tgcacccacg gagcctgtca
     2041 gtggccccaa agtggagcgg cggccttcca ggaaaccacc cacgtcccct tcgagcccgc
     2101 cagcgccagt acctcgagtt ctcgctgccc cgcagaactc acctgtggga ccagggttgt
     2161 caatttccca gctggcggcc tccccgcggt ccccgactca gcactgcttg gccaggccta
     2221 cttcacagct accccatggc tctcaggcct ccccggccca ggcacaggag ttcccgttgg
     2281 aggccggtat ctcccacctg gaagccgacc tgagccagac ctccctggtc ctggaaacat
     2341 ccattgccga acagttacag gagctgccgt tcacgccttt gcatgcccct attgttgtgg
     2401 gaacccagac caggagctct gcagggcagc cctcgagagc ctccatggtg ctcctgcagt
     2461 cctccggctt tcccgagatt ctggatgcca ataaacagcc agccgaggct gtcagcgcta
     2521 cagaacctgt gacgtttaac cctcagaagg aagaatcaga ttgtctacaa agcaacgaga
     2581 tggtgctaca gtttcttgcc tttagcagag tggcccagga ctgccgagga acatcatggc
     2641 caaagactgt gtatttcacc ttccagttct accgcttccc acccgcaacg acgccacgac
     2701 tgcagctggt ccagctggat gaggccggcc agcccagctc tggcgccctg acccacatcc
     2761 tcgtgcctgt gagcagagat ggcacctttg atgctgggtc tcctggcttc cagctgaggt
     2821 acatggtggg ccctgggttc ctgaagccag gtgagcggcg ctgctttgcc cgctacctgg
     2881 ccgtgcagac cctgcagatt gacgtctggg acggagactc cctgctgctc atcggatctg
     2941 ctgccgtcca gatgaagcat ctcctccgcc aaggccggcc ggctgtgcag gcctcccacg
     3001 agcttgaggt cgtggcaact gaatacgagc aggacaacat ggtggtgagt ggagacatgc
     3061 tggggtttgg ccgcgtcaag cccatcggcg tccactcggt ggtgaagggc cggctgcacc
     3121 tgactttggc caacgtgggt cacccgtgtg aacagaaagt gagaggttgt aggatatcgg
     3181 taccagtcga ctctagagga tcccgggtgg catccctgtg acccctcccc agtgcctctc
     3241 ctggccctgg aagttgccac tccagtgccc accagccttg tcctaataaa attaagttgc
     3301 atcattttgt ctgactaggt gtccttctat aatattatgg ggtggagggg ggtggtatgg
     3361 agcaaggggc aagttgggaa gacaacctgt agggcctgcg gggtctattg ggaaccaagc
     3421 tggagtgcag tggcacaatc ttggctcact gcaatctccg cctcctgggt tcaagcgatt
     3481 ctcctgcctc agcctcccga gttgttggga ttccaggcat gcatgaccag gctcagctaa
     3541 tttttgtttt tttggtagag acggggtttc accatattgg ccaggctggt ctccaactcc
     3601 taatctcagg tgatctaccc accttggcct cccaaattgc tgggattaca ggcgtgaacc
     3661 actgctccct tccctgtcct tctgatttta aaataactat accagcagga ggacgtccag
     3721 acacagcata ggctacctgg ccatgcccaa ccggtgggac atttgagttg cttgcttggc
     3781 actgtcctct catgcgttgg gtccactcag tagatgcctg ttgaattggg tacgcggcca
     3841 gcttggctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     3901 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     3961 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     4021 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     4081 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctcggcctc tgagctattc
     4141 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cctcgaggaa
     4201 ctgaaaaacc agaaagttaa ttccctatag tgagtcgtat taaattcgta atcatgtcat
     4261 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     4321 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     4381 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     4441 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     4501 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     4561 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     4621 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     4681 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     4741 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     4801 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     4861 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     4921 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     4981 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     5041 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     5101 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     5161 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     5221 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     5281 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     5341 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     5401 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     5461 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     5521 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     5581 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     5641 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     5701 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     5761 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     5821 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     5881 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     5941 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     6001 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     6061 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     6121 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     6181 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     6241 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     6301 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     6361 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     6421 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     6481 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     6541 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     6601 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     6661 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     6721 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     6781 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     6841 acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
     6901 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     6961 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     7021 tgccaagctg atctatacat tgaatcaata ttggcaatta gccatattag tcattggtta
     7081 tatagcataa atcaatattg gctattggcc attgcatacg ttgtatctat atcataatat
     7141 gtacatttat attggctcat gtccaatatg accgccatgt t
//
