LOCUS       sequence-8494-ex        5010 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5010)
  AUTHORS   Hsiao EC, Boudignon BM, Chang WC, Bencsik M, Peng J, Nguyen TD, 
            Manalac C, Halloran BP, Conklin BR, Nissenson RA
  TITLE     Osteoblast expression of an engineered Gs-coupled receptor 
            dramatically increases bone mass.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Jan 29. 105(4):1209-14.
  PUBMED    18212126
REFERENCE   2  (bases 1 to 5010)
  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: "Proc Natl 
            Acad Sci U S A. 2008 Jan 29. 105(4):1209-14."
COMMENT     Sequence Label: pUHG10.3-TetO-Rs1
FEATURES             Location/Qualifiers
     source          1..5010
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    join(15..33,34..56,57..75,76..98,99..117,118..140,141..159,
                     160..182,183..201,202..224,225..243,244..266,267..285)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        318..355
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     352..376
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          1801..2373
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2505..2524)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2570..2625
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2624..2643)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3016..3033)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3187..3775)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3267..3286)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3946..4737,4738..4806))
                     /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     4569..4588
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4807..4911)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4979..4997
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ctcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc
       61 tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg atagagaaaa
      121 gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt cgagtttacc
      181 actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta tcagtgatag
      241 agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag
      301 ctcggtaccc gggtcgagta ggcgtgtacg gtgggaggcc tatataagca gagctcgttt
      361 agtgaaccgt cagatcgcct ggagacgcca tccacgctgt tttgacctcc atagaagaca
      421 ccgggaccga tccagcctcc gcggccccga attcgccctt gatatcacca tgaagacgat
      481 catcgccctg agctacatct tctgcctggt attcgccgac tacaaggacg atgatgacgc
      541 cagcggccgc gcccgggcgg atcccccggg ctgcaggatg gacaaacttg atgctaatgt
      601 gagttctgag gagggtttcg ggtcagtgga gaaggtggtg ctgctcacgt ttctctcgac
      661 ggttatcctg atggccatct tggggaacct gctggtgatg gtggctgtgt gctgggacag
      721 gcagctcagg aaaataaaaa caaattattt cattgtatct cttgcttttg cggatctgct
      781 ggtttcggtg ctggtgatgc cctttggtgc cattgagctg gttcaagaca tctggattta
      841 tggggaggtg ttttgtcttg ttcggacatc tctggccgtc ctgctcacaa cggcatcgat
      901 ttttcacctg tgctgcattt ctctggatag gtattacgcc atctgctgcc agcctttggt
      961 ctataggaac aagatgaccc ctctgcgcat cgcattaatg ctgggaggct gctgggtcat
     1021 ccccacgttt atttcttttc tccctataat gcaaggctgg aataacattg gcataattga
     1081 tttgatagaa aagaggaagt tcaaccagaa ctctaactct acgtactgtg tcttcatggt
     1141 caacaagccc tacgccatca cctgctctgt ggtggccttc tacatcccat ttctcctcat
     1201 ggtgctggcc tattaccgca tctatgtcac agctaaggag catgcccatc agatccagat
     1261 gttacaacgg gcaggagcct cctccgagag caggcctcag tcggcagacc agcatagcac
     1321 tcatcgcatg aggacagaga ccaaagcagc caagacccta tgcatcatca tgggttgctt
     1381 ctgcctctgc tgggcaccat tctttgtcac caatattgtg gatcctttca tagactacac
     1441 tgtccctggg caggtgtgga ctgctttcct ctggctcggc tatatcaatt ccgggttgaa
     1501 cccttttctc tacgccttct tgaataagtc ttttagacgt gccttcctca tcatcctctg
     1561 ctgtgatgat gagcgctacc gaagaccttc cattctgggc cagactgtcc cttgttcaac
     1621 cacaaccatt aatggatcca cacatgtact aagggatgca gtggagtgtg gtggccagtg
     1681 ggagagtcag tgtcacccgc cagcaacttc tcctttggtg gctgctcagc ccagtgacac
     1741 ttaggcggcc gcgaattcga gctcggtacc cggggatcct ctagagctga gaacttcagg
     1801 gtgagtttgg ggacccttga ttgttctttc tttttcgcta ttgaaaaatt catgttatat
     1861 ggagggggca aagttttcag ggtgttgttt agaatgggaa gatgtccctt gtatcaccat
     1921 ggaccctcat gataattttg tttctttcac tttctactct gttgacaacc attgtctcct
     1981 cttattttct tttcattttc tgtaactttt ttcgttaaac tttagcttgc atttgtaacg
     2041 aatttttaaa ttcactttcg tttatttgtc agattgtaag tactttctct aatcactttt
     2101 ttttcaaggc aatcagggta attatattgt acttcagcac agttttagag aacaattgtt
     2161 ataattaaat gataaggtag aatatttctg catataaatt ctggctggcg tggaaatatt
     2221 cttattggta gaaacaacta catcctggta atcatcctgc ctttctcttt atggttacaa
     2281 tgatatacac tgtttgagat gaggataaaa tactctgagt ccaaaccggg cccctctgct
     2341 aaccatgttc atgccttctt ctttttccta cagctcctgg gcaacgtgct ggttgttgtg
     2401 ctgtctcatc attttggcaa agaattcact cctcaggtgc aggctgccta tcagaaggtg
     2461 gtggctggtg tggccaatgc cctggctcac aaataccact gagatctttt tccctctgcc
     2521 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctgggta ataaaggaaa
     2581 tttattttca ttgcaatagt gtgtgggaat tttttgtgtc tctcactcgg aaggacatat
     2641 gggagggcaa atcatttaaa acatcagaat gagtatttgg tttagagttt ggcaacatat
     2701 gccatatgct ggctgccatg aacaaaggtg gctataaaga ggtcatcagt atatgaaaca
     2761 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     2821 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     2881 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     2941 atgaactcga ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg
     3001 gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc
     3061 ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg
     3121 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct
     3181 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca
     3241 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct
     3301 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc
     3361 gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat ctcagttcgg tgtaggtcgt
     3421 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc
     3481 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc
     3541 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg
     3601 gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc tgctgaagcc
     3661 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag
     3721 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga
     3781 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat
     3841 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag
     3901 ttttaaatca atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat
     3961 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     4021 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat
     4081 accgcgagac ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag
     4141 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     4201 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc
     4261 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     4321 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     4381 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     4441 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     4501 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     4561 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     4621 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     4681 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     4741 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     4801 actcatactc ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag
     4861 cggatacata tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc
     4921 ccgaaaagtg ccacctgacg tctaagaaac cattattatc atgacattaa cctataaaaa
     4981 taggcgtatc acgaggccct ttcgtcttca
//
