LOCUS       sequence-178204-        4418 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  reduced the expression of gamma-tubulin in human cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4418)
  AUTHORS   Alvarado-Kristensson M, Rodriguez MJ, Silio V, Valpuesta JM, Carrera
            AC
  TITLE     SADB phosphorylation of gamma-tubulin regulates centrosome 
            duplication.
  JOURNAL   Nat Cell Biol. 2009 Sep;11(9):1081-92. doi: 10.1038/ncb1921. Epub 
            2009 Aug 2.
  PUBMED    19648910
REFERENCE   2  (bases 1 to 4418)
  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"; doi: "10.1038/ncb1921"; 
            journalName: "Nat Cell Biol"; date: "2009-09"; volume: "11"; issue: 
            "9"; pages: "1081-92"
COMMENT     Sequence Label: pTER GAMMA-TUBULIN shRNA Human
FEATURES             Location/Qualifiers
     source          1..4418
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     161..180
                     /label=H1
                     /note="Human H1 promoter, forward primer"
     protein_bind    226..244
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     complement(416..433)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    422..646
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      692..1120
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(779..798)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     989..1010
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1129..1149)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1134..1463
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1314..1449
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1376..1395
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1511..1558
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1577..1951
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    2081..2202
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2118..2137)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2172..2191
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2251..2267)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2251..2267)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2264..2286)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2275..2291
                     /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(2299..2305,2306..2323,2324..2329))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2344..2365
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2482..2499)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2653..3241)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2733..2752)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3412..4203,4204..4272))
                     /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     4035..4054
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4273..4377)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gagatccccc gggctgcagg aattcgaacg ctgacgtcat caacccgctc caaggaatcg
       61 cgggcccagt gtcactaggc gggaacaccc agcgcgcgtg cgccctggca ggaagatggc
      121 tgtgagggac aggggagtgg cgccctgcaa tatttgcatg tcgctatgtg ttctgggaaa
      181 tcaccataaa cgtgaaatgt ctttggattt gggaatctta taagttccct atcagtgata
      241 gagatcccgg aggacatgtt caaggacttc aagagagtcc ttgaacatgt cctccttttt
      301 ggaaaagctt ggtaccgagc tcggatccac tagtccagtg tggtggaatt ctgcagatat
      361 ccagcacagt ggcggccgct cgagtctaga gggcccgttt aaacccgctg atcagcctcg
      421 actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc ttccttgacc
      481 ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc atcgcattgt
      541 ctgagtaggt gtcattctat tctggggggt ggggtggggc aggacagcaa gggggaggat
      601 tgggaagaca atagcaggca tgctggggat gcggtgggct ctatggcttc tgaggcggaa
      661 agaaccagct ggggctctag ggggtatccc cacgcgccct gtagcggcgc attaagcgcg
      721 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct agcgcccgct
      781 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
      841 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
      901 cttgattagg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
      961 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     1021 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1081 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc
     1141 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     1201 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     1261 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     1321 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     1381 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     1441 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcggatctg
     1501 atcagcacgt gttgacaatt aatcatcggc atagtatatc ggcatagtat aatacgacaa
     1561 ggtgaggaac taaaccatgg ccaagttgac cagtgccgtt ccggtgctca ccgcgcgcga
     1621 cgtcgccgga gcggtcgagt tctggaccga ccggctcggg ttctcccggg acttcgtgga
     1681 ggacgacttc gccggtgtgg tccgggacga cgtgaccctg ttcatcagcg cggtccagga
     1741 ccaggtggtg ccggacaaca ccctggcctg ggtgtgggtg cgcggcctgg acgagctgta
     1801 cgccgagtgg tcggaggtcg tgtccacgaa cttccgggac gcctccgggc cggccatgac
     1861 cgagatcggc gagcagccgt gggggcggga gttcgccctg cgcgacccgg ccggcaactg
     1921 cgtgcacttc gtggccgagg agcaggactg acacgtgcta cgagatttcg attccaccgc
     1981 cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct ggatgatcct
     2041 ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta ttgcagctta
     2101 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     2161 gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct gtataccgtc
     2221 gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta
     2281 tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc
     2341 ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg
     2401 aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg
     2461 tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     2521 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     2581 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     2641 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     2701 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     2761 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     2821 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     2881 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     2941 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     3001 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     3061 gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct
     3121 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     3181 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     3241 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     3301 agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa
     3361 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     3421 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     3481 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     3541 aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc
     3601 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     3661 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     3721 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     3781 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     3841 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     3901 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     3961 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     4021 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     4081 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     4141 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     4201 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     4261 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     4321 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4381 atttccccga aaagtgccac ctgacgtcga cggatcgg
//
