LOCUS       sequence-178305-        6609 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFLAG-CMV2 with cDNA encoding HsNPHP4 aa residues 1-649.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6609)
  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 6609)
  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: LP454
FEATURES             Location/Qualifiers
     source          1..6609
                     /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    2633..3255
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(2832..2851)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3279..3299)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3284..3613
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3464..3599
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(3651..3670)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(3652..3670)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3684..3700)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3684..3700)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3697..3719)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3708..3724
                     /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(3732..3738,3739..3756,3757..3762))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3777..3798
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3915..3932)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4086..4674)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4166..4185)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4845..5636,5637..5705))
                     /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     5468..5487
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5706..5810)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5837..6292
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5924..5943)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6134..6155
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6418..6440
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6432..6449
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6433..6449
                     /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 gatatcggta ccagtcgact ctagaggatc ccgggtggca
     2641 tccctgtgac ccctccccag tgcctctcct ggccctggaa gttgccactc cagtgcccac
     2701 cagccttgtc ctaataaaat taagttgcat cattttgtct gactaggtgt ccttctataa
     2761 tattatgggg tggagggggg tggtatggag caaggggcaa gttgggaaga caacctgtag
     2821 ggcctgcggg gtctattggg aaccaagctg gagtgcagtg gcacaatctt ggctcactgc
     2881 aatctccgcc tcctgggttc aagcgattct cctgcctcag cctcccgagt tgttgggatt
     2941 ccaggcatgc atgaccaggc tcagctaatt tttgtttttt tggtagagac ggggtttcac
     3001 catattggcc aggctggtct ccaactccta atctcaggtg atctacccac cttggcctcc
     3061 caaattgctg ggattacagg cgtgaaccac tgctcccttc cctgtccttc tgattttaaa
     3121 ataactatac cagcaggagg acgtccagac acagcatagg ctacctggcc atgcccaacc
     3181 ggtgggacat ttgagttgct tgcttggcac tgtcctctca tgcgttgggt ccactcagta
     3241 gatgcctgtt gaattgggta cgcggccagc ttggctgtgg aatgtgtgtc agttagggtg
     3301 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3361 agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3421 tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc
     3481 gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc
     3541 cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct
     3601 aggcttttgc aaaaagctcc tcgaggaact gaaaaaccag aaagttaatt ccctatagtg
     3661 agtcgtatta aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc
     3721 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     3781 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3841 tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg
     3901 cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     3961 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     4021 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     4081 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     4141 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     4201 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     4261 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     4321 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     4381 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     4441 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     4501 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     4561 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     4621 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     4681 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     4741 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     4801 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     4861 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     4921 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     4981 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     5041 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     5101 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     5161 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     5221 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     5281 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     5341 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     5401 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     5461 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     5521 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     5581 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     5641 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     5701 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     5761 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     5821 cgaaaagtgc cacctgacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt
     5881 acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc
     5941 ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct
     6001 ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat
     6061 ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc
     6121 acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc
     6181 tattcttttg atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg
     6241 atttaacaaa aatttaacgc gaattttaac aaaatattaa cgcttacaat ttccattcgc
     6301 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     6361 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     6421 agtcacgacg ttgtaaaacg acggccagtg ccaagctgat ctatacattg aatcaatatt
     6481 ggcaattagc catattagtc attggttata tagcataaat caatattggc tattggccat
     6541 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     6601 cgccatgtt
//
