LOCUS       sequence-177051-        5843 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Labels mother centrioles/basal bodies.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5843)
  TITLE     GFP-hChibby1
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5843)
  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: 
            "Unpublished"
COMMENT     Sequence Label: GFP-hChibby1
FEATURES             Location/Qualifiers
     source          1..5843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        27..1004
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1039..1057
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1039..1056
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             1092..1805
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1122..1150)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1745..1765
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2875..2894)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2876..2894)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     polyA_signal    2899..3033
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2923..2942)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2977..2996
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3146..3166)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3146..3164)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3185..3201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3185..3201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3198..3220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3209..3225
                     /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(3233..3239,3240..3257,3258..3263))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3278..3299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3416..3433)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3587..4175)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3667..3686)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4346..5137,5138..5206))
                     /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     4969..4988
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5207..5311)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(5337..5792)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5474..5495)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5686..5705
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 cctcttcgct attacgccag tcgaccgcca attcaatatg gcgtatatgg actcatgcca
       61 attcaatatg gtggatctgg acctgtgcca attcaatatg gcgtatatgg actcgtgcca
      121 attcaatatg gtggatctgg accccagcca attcaatatg gcggacttgg caccatgcca
      181 attcaatatg gcggacctgg cactgtgcca actggggagg ggtctacttg gcacggtgcc
      241 aagtttgagg aggggtcttg gccctgtgcc aagtccgcca tattgaattg gcatggtgcc
      301 aataatggcg gccatattgg ctatatgcca ggatcaatat ataggcaata tccaatatgg
      361 ccctatgcca atatggctat tggccaggtt caatactatg tattggccct atgccatata
      421 gtattccata tatgggtttt cctattgacg tagatagccc ctcccaatgg gcggtcccat
      481 ataccatata tggggcttcc taataccgcc catagccact cccccattga cgtcaatggt
      541 ctctatatat ggtctttcct attgacgtca tatgggcggt cctattgacg tatatggcgc
      601 ctcccccatt gacgtcaatt acggtaaatg gcccgcctgg ctcaatgccc attgacgtca
      661 ataggaccac ccaccattga cgtcaatggg atggctcatt gcccattcat atccgttctc
      721 acgcccccta ttgacgtcaa tgacggtaaa tggcccactt ggcagtacat caatatctat
      781 taatagtaac ttggcaagta cattactatt ggaagtacgc cagggtacat tggcagtact
      841 cccattgacg tcaatggcgg taaatggccc gcgatggctg ccaagtacat ccccattgac
      901 gtcaatgggg aggggcaatg acgcaaatgg gcgttccatt gacgtaaatg ggcggtaggc
      961 gtgcctaatg ggaggtctat ataagcaatg ctcgtttagg gaaccgccat tctgcctggg
     1021 gacgtcggag caagcttgat ttaggtgaca ctatagaata caagctactt gttctttttg
     1081 caggatctac catgagtaaa ggagaagaac ttttcactgg agttgtccca attcttgttg
     1141 aattagatgg tgatgttaat gggcacaaat tttctgtcag tggagagggt gaaggtgatg
     1201 caacatacgg aaaacttacc cttaaattta tttgcactac tggaaaacta cctgttccat
     1261 ggccaacact tgtcactact ctcacttatg gtgttcaatg cttttcaaga tacccagatc
     1321 atatgaaacg gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
     1381 ctatattttt caaagatgac gggaactaca agacacgtgc tgaagtcaag tttgaaggtg
     1441 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
     1501 ttggacacaa attggaatac aactataact cacacaatgt atacatcatg gcagacaaac
     1561 aaaagaatgg aatcaaagtt aacttcaaaa ttagacacaa cattgaagat ggaagcgttc
     1621 aactagcaga ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag
     1681 acaaccatta cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc
     1741 acatggtcct tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat
     1801 acaaaggatc ccatcgattc gaattcatgc ctttctttgg gaatacgttc agtccgaaga
     1861 agacacctcc tcggaagtcg gcatctctct ccaacctgca ttctttggat cgatcaaccc
     1921 gggaggtgga gctgggcttg gaatacggat ccccgactat gaacctggca gggcaaagcc
     1981 tgaagtttga aaatggccag tggatagcag agacaggggt tagtggcggt gtggaccgga
     2041 gggaggttca gcgccttcgc aggcggaacc agcagttgga ggaagagaac aatctcttgc
     2101 ggctgaaagt ggacatctta ttagacatgc tttcagagtc cactgccgaa tcccacttaa
     2161 tggagaagga actggatgaa ctgaggatca gccggaagag aaaatgaaga ccccagagac
     2221 atttattggg gagtaggatg tggctgagtg cttttttttt tggccagact agcggattca
     2281 gtcctggaag agagtatcat ataatgagac ccacaggcac tggcaccctt gggttggcaa
     2341 tagaaggtga catggaatgg agaaaaccaa gattccagat ggggatagta actagaaggt
     2401 gcttcagatg cactgcctgc gggtgccagt ctgaaaacca gaccgcacag aggcctgggg
     2461 ctgctgatga gcttttcggt gctctccaca cacaagctcg caaacacaca tgtcccagaa
     2521 tagctctgtt gggttgtgtt gggagaagcg gctggagttc attctctcac ccccttatgt
     2581 tggtgtttgg cgtgtgacag cagttctaca gagctctgtg ttggggtcat ggatgagcgg
     2641 ctctcttggc tcttaaaggc aggcctctct cttcttgcct ttaaagaatc ctccttcctc
     2701 acacctggcc tcctctggct tcagcttctc agcagcaagc accagccttc cacaacaaca
     2761 ctatattttt atgctacttt cctgtttgca ctactacttt tttattaaac gatgttaaat
     2821 aaaaaaaaaa aaaaaaaaaa aaaaaaaaaa rwmwmgmgcc tctagattct gcagccctat
     2881 agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc
     2941 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     3001 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     3061 tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgcggc gccaatgcat
     3121 tgggcccggt acccagcttt tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat
     3181 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     3241 gagccgggag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     3301 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     3361 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     3421 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3481 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3541 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     3601 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     3661 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     3721 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     3781 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     3841 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     3901 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     3961 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     4021 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4081 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4141 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4201 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     4261 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     4321 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     4381 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     4441 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     4501 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     4561 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     4621 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     4681 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     4741 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     4801 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     4861 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     4921 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     4981 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     5041 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     5101 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     5161 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     5221 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     5281 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat
     5341 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     5401 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     5461 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     5521 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     5581 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     5641 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     5701 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     5761 cgccgcgctt aatgcgccgc tacagggcgc gtcccattcg ccattcaggc tgcgcaactg
     5821 ttgggaaggg cgatcggtgc ggg
//
