LOCUS       sequence-9329-ex        3341 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 3341)
  AUTHORS   Deans TL, Cantor CR, Collins JJ
  TITLE     A tunable genetic switch based on RNAi and repressor proteins for 
            regulating gene expression in mammalian cells.
  JOURNAL   Cell. 2007 Jul 27. 130(2):363-72.
  PUBMED    17662949
REFERENCE   2  (bases 1 to 3341)
  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: "Cell. 2007 
            Jul 27. 130(2):363-72."
COMMENT     Sequence Label: TLD_062
FEATURES             Location/Qualifiers
     source          1..3341
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        11..29
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     11..28
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     175..195
                     /label=hU6-F
                     /note="Human U6 promoter, forward primer"
     primer_bind     346..365
                     /label=LKO.1 5'
                     /note="Human U6 promoter, forward primer"
     protein_bind    377..395
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O1 for the tetR and tetA genes"
     primer_bind     complement(489..508)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(490..508)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(515..532)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(515..531)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(524..546)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      672..1127
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(759..778)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     969..990
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1208..1312
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1313..1381,1382..2173)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMPTTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1531..1550)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2344..2932
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2833..2852
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3086..3103
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3220..3241
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3256..3261,3262..3279,3280..3286)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3294..3310
                     /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)."
     primer_bind     3299..3321
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3318..3334
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3318..3334
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 acgccaagct atttaggtga cactatagaa tactcaagct atgcatccaa cgcgttggga
       61 gctctcccat atggtcgacc tgcaggcggc cgcgaattca ctagtgatta ggcaaaacgc
      121 accacgtgac ggagcgtgac cgcgcgccga gcgcgcgcca aggtcgggca ggaagagggc
      181 ctatttccca tgattccttc atatttgcat atacgataca aggctgttag agagataatt
      241 agaattaatt tgactgtaaa cacaaagata ttagtacaaa atacgtgacg tagaaagtaa
      301 taatttcttg ggtagtttgc agttttaaaa ttatgtttta aaatggacta tcatatgctt
      361 accgtaactt gaaagtactc tatcattgat agagttatat atcttgtgga aaggacgaaa
      421 caccgctaca caaatcagcg atttttcaag agaaaatcgc tgatttgtgt agaaaaaggc
      481 ccaattcgcc ctatagtgag tcgtattaca attcactggc cgtcgtttta caacgtcgtg
      541 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
      601 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
      661 atggcgaatg gacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
      721 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
      781 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
      841 gttccgattt agagctttac ggcacctcga ccgcaaaaaa cttgatttgg gtgatggttc
      901 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
      961 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     1021 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     1081 acaaatattt aacgcgaatt ttaacaaaat attaacgttt acaatttcgc ctgatgcggt
     1141 attttctcct tacgcatctg tgcggtattt cacaccgcat acaggtggca cttttcgggg
     1201 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     1261 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1321 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1381 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1441 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1501 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     1561 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1621 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1681 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1741 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1801 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     1861 aatgccaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     1921 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     1981 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2041 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2101 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2161 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2221 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2281 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2341 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2401 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2461 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2521 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2581 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2641 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2701 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2761 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2821 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2881 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     2941 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3001 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3061 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3121 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3181 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     3241 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     3301 cggataacaa tttcacacag gaaacagcta tgaccatgat t
//
