LOCUS       sequence-178306-        6999 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pFLAG-CMV2 with cDNA encoding HsNPHP4 aa residues 649-1426.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6999)
  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 6999)
  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: LP455
FEATURES             Location/Qualifiers
     source          1..6999
                     /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    3023..3645
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3222..3241)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     primer_bind     complement(3669..3689)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3674..4003
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3854..3989
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(4041..4060)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4042..4060)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4074..4090)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4074..4090)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4087..4109)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4098..4114
                     /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(4122..4128,4129..4146,4147..4152))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4167..4188
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4305..4322)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4476..5064)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4556..4575)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5235..6026,6027..6095))
                     /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     5858..5877
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6096..6200)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      6227..6682
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6314..6333)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6524..6545
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6808..6830
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6822..6839
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6823..6839
                     /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 gcagccttta
      661 gcagagtggc ccaggactgc cgaggaacat catggccaaa gactgtgtat ttcaccttcc
      721 agttctaccg cttcccaccc gcaacgacgc cacgactgca gctggtccag ctggatgagg
      781 ccggccagcc cagctctggc gccctgaccc acatcctcgt gcctgtgagc agagatggca
      841 cctttgatgc tgggtctcct ggcttccagc tgaggtacat ggtgggccct gggttcctga
      901 agccaggtga gcggcgctgc tttgcccgct acctggccgt gcagaccctg cagattgacg
      961 tctgggacgg agactccctg ctgctcatcg gatctgctgc cgtccagatg aagcatctcc
     1021 tccgccaagg ccggccggct gtgcaggcct cccacgagct tgaggtcgtg gcaactgaat
     1081 acgagcagga caacatggtg gtgagtggag acatgctggg gtttggccgc gtcaagccca
     1141 tcggcgtcca ctcggtggtg aagggccggc tgcacctgac tttggccaac gtgggtcacc
     1201 cgtgtgaaca gaaagtgaga ggttgtagca cattgccacc gtccagatct cgggtcatct
     1261 caaacgatgg agccagccgc ttctctggag gcagcctcct cacgactgga agctcaaggc
     1321 gaaaacacgt ggtgcaagca cagaagctgg cggacgtgga cagtgagctg gctgccatgc
     1381 tactgaccca tgcccggcag ggcaaggggc cccaggacgt cagccgcgag tcggatgcca
     1441 cccgcaggcg taagctggag cggatgaggt ctgtgcgcct gcaggaggcc gggggagact
     1501 tgggccggcg tgggacgagc gtgttggcgc agcagagcgt ccgcacacag cacttgcggg
     1561 acctacaggt catcgccgcc taccgggaac gcacgaaggc cgagagcatc gccagcctgc
     1621 tgagcctggc catcaccacg gagcacacgc tccacgccac gctgggggtc gccgagttct
     1681 ttgagtttgt gcttaagaac ccccacaaca cacagcacac ggtgactgtg gagatcgaca
     1741 accccgagct cagcgtcatc gtggacagtc aggagtggag ggacttcaag ggtgctgctg
     1801 gcctgcacac accggtggag gaggacatgt tccacctgcg tggcagcctg gccccccagc
     1861 tctacctgcg cccccacgag accgcccacg tccccttcaa gttccagagc ttctctgcag
     1921 ggcagctggc catggtgcag gcctctcctg ggttgagcaa cgagaagggc atggacgccg
     1981 tgtcaccttg gaagtccagc gcagtgccca ctaaacacgc caaggtcttg ttccgagcga
     2041 gtggtggcaa gcccatcgcc gtgctctgcc tgactgtgga gctgcagccc cacgtggtgg
     2101 accaggtctt ccgcttctat cacccggagc tctccttcct gaagaaggcc atccgcctgc
     2161 cgccctggca cacatttcca ggtgctccgg tgggaatgct tggtgaggac cccccagtcc
     2221 atgttcgctg cagcgacccg aacgtcatct gtgagaccca gaatgtgggc cccggggaac
     2281 cacgggacat atttctgaag gtggccagtg gtccaagccc ggagatcaaa gacttctttg
     2341 tcatcattta ctcggatcgt tggctggcga cacccacaca gacgtggcag gtctacctcc
     2401 actccctgca gcgcgtggat gtctcctgcg tcgcaggcca gctgacccgc ctgtcccttg
     2461 tccttcgggg gacacagaca gtgaggaaag tgagagcttt cacctctcat ccacaggagc
     2521 tgaagacaga ccccaaaggt gtcttcgtgc tgccgcctcg tggggtgcag gacctgcatg
     2581 ttggcgtgag gccccttagg gccggcagcc gctttgtcca tctcaacctg gtggacgtgg
     2641 attgccacca gctggtggcc tcctggctcg tgtgcctctg ctgccgccag ccgctcatct
     2701 ccaaggcctt tgagatcatg ttggctgcgg gcgaagggaa gggtgtcaac aagaggatca
     2761 cctacaccaa cccctacccc tcccggagga cattccacct gcacagcgac cacccggagc
     2821 tgctgcggtt cagagaggac tccttccagg tcgggggtgg agagacctac accatcggct
     2881 tgcagtttgc gcctagtcag agagtgggtg aggaggagat cctgatctac atcaatgacc
     2941 atgaggacaa aaacgaagag gcattttgcg tgaaggtcat ctaccagtga gatatcggta
     3001 ccagtcgact ctagaggatc ccgggtggca tccctgtgac ccctccccag tgcctctcct
     3061 ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat taagttgcat
     3121 cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg tggtatggag
     3181 caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg aaccaagctg
     3241 gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc aagcgattct
     3301 cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc tcagctaatt
     3361 tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct ccaactccta
     3421 atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg cgtgaaccac
     3481 tgctcccttc cctgtccttc tgattttaaa ataactatac cagcaggagg acgtccagac
     3541 acagcatagg ctacctggcc atgcccaacc ggtgggacat ttgagttgct tgcttggcac
     3601 tgtcctctca tgcgttgggt ccactcagta gatgcctgtt gaattgggta cgcggccagc
     3661 ttggctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc cagcaggcag
     3721 aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt ccccaggctc
     3781 cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca tagtcccgcc
     3841 cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg
     3901 ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca
     3961 gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc tcgaggaact
     4021 gaaaaaccag aaagttaatt ccctatagtg agtcgtatta aattcgtaat catgtcatag
     4081 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     4141 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     4201 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     4261 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     4321 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4381 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4441 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4501 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4561 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4621 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4681 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4741 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4801 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4861 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4921 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4981 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     5041 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5101 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5161 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5221 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5281 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5341 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5401 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5461 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5521 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5581 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5641 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5701 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5761 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5821 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5881 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5941 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     6001 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6061 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6121 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6181 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgc gccctgtagc
     6241 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     6301 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     6361 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     6421 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     6481 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     6541 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     6601 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac
     6661 aaaatattaa cgcttacaat ttccattcgc cattcaggct gcgcaactgt tgggaagggc
     6721 gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc
     6781 gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg
     6841 ccaagctgat ctatacattg aatcaatatt ggcaattagc catattagtc attggttata
     6901 tagcataaat caatattggc tattggccat tgcatacgtt gtatctatat cataatatgt
     6961 acatttatat tggctcatgt ccaatatgac cgccatgtt
//
