LOCUS       sequence-9924-ex        5392 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 5392)
  AUTHORS   Mon H, Sugahara R, Hong SM, Lee JM, Kamachi Y, Kawaguchi Y, Kusakabe
            T
  TITLE     Analysis of protein interactions with two-hybrid system in cultured 
            insect cells.
  JOURNAL   Anal Biochem. 2009 Sep 15;392(2):180-2. doi: 
            10.1016/j.ab.2009.05.033.
  PUBMED    19481053
REFERENCE   2  (bases 1 to 5392)
  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: 
            "10.1016/j.ab.2009.05"; journalName: "Anal Biochem"; date: 
            "2009-09-15- 15"; volume: "392"; issue: "2"; pages: "180-2"
COMMENT     Sequence Label: pUAS-Luc
FEATURES             Location/Qualifiers
     source          1..5392
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..155
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     147..164
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     148..164
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     174..193
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        174..192
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     217..233
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     218..234
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(268..284)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(268..284)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    544..568
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             625..2277
                     /codon_start=1
                     /gene="luc"
                     /product="firefly luciferase"
                     /label=luciferase
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     primer_bind     complement(691..709)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     protein_bind    complement(2350..2374)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     primer_bind     complement(2409..2428)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2463..2482
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2538..2558)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(2538..2556)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2577..2593)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2577..2593)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2590..2612)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2601..2617
                     /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(2625..2631,2632..2649,2650..2655))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2670..2691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2808..2825)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2979..3567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3059..3078)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(3749..4282)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(join(4471..4476,4477..4493,4494..4499))
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     rep_origin      4944..5372
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5031..5050)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5241..5262
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
       61 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
      121 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcgtaatacg
      181 actcactata gggcgaattg ggtaccgggc cccccctcga ggtcgacggt atcgataagc
      241 ttgatatcga attcctgcag cccgggggat ccactagttc tagagcggcc gcccgctaga
      301 cggagtactg tcctccgagc ggagtactgt cctccgagcg gagtactgtc ctccgagcgg
      361 agtactgtcc tccgatcacg ttcattctgt ttaaatatgc gacaccgctg gcgggaagca
      421 gaattatttg aaacgcaata cgaagcagac gaagagagtt cagtgcaaac tgaaaacaag
      481 tttttcgacg atttatttat tgtttattgt gaagaagtat ctacagtgat taatattcaa
      541 aatacaagtt tgtacaaaaa agcaggcttc gaaggagata gaaccaattc tctaaggaaa
      601 tacttaacca tggtcgagat ccaaatggaa gacgccaaaa acataaagaa aggcccggcg
      661 ccattctatc ctctagagga tggaaccgct ggagagcaac tgcataaggc tatgaagaga
      721 tacgccctgg ttcctggaac aattgctttt acagatgcac atatcgaggt gaacatcacg
      781 tacgcggaat acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg
      841 aatacaaatc acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg
      901 ttgggcgcgt tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt
      961 gaattgctca acagtatgaa catttcgcag cctaccgtag tgtttgtttc caaaaagggg
     1021 ttgcaaaaaa ttttgaacgt gcaaaaaaaa ttaccaataa tccagaaaat tattatcatg
     1081 gattctaaaa cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta
     1141 cctcccggtt ttaatgaata cgattttgta ccagagtcct ttgatcgtga caaaacaatt
     1201 gcactgataa tgaattcctc tggatctact gggttaccta agggtgtggc ccttccgcat
     1261 agaactgcct gcgtcagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt
     1321 ccggatactg cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca
     1381 ctcggatatt tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg
     1441 tttttacgat cccttcagga ttacaaaatt caaagtgcgt tgctagtacc aaccctattt
     1501 tcattcttcg ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt
     1561 gcttctgggg gcgcacctct ttcgaaagaa gtcggggaag cggttgcaaa acgcttccat
     1621 cttccaggga tacgacaagg atatgggctc actgagacta catcagctat tctgattaca
     1681 cccgaggggg atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag
     1741 gttgtggatc tggataccgg gaaaacgctg ggcgttaatc agagaggcga attatgtgtc
     1801 agaggaccta tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt
     1861 gacaaggatg gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc
     1921 ttcatagttg accgcttgaa gtctttaatt aaatacaaag gatatcaggt ggcccccgct
     1981 gaattggaat cgatattgtt acaacacccc aacatcttcg acgcgggcgt ggcaggtctt
     2041 cccgacgatg acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg
     2101 atgacggaaa aagagatcgt ggattacgtg gccagtcaag taacaaccgc gaaaaagttg
     2161 cgcggaggag ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca
     2221 agaaaaatca gagagatcct cataaaggcc aagaagggcg gaaagtccaa attgtaaaat
     2281 gtaactgtat tcagcgatga cgaaattctt agctattgta atcctccgag gcctcgaggt
     2341 cgaatctaga cccagctttc ttgtacaaag tggtttgttt attgcagctt ataatggtta
     2401 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2461 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgcggccgcc accgcggtgg
     2521 agctccagct tttgttccct ttagtgaggg ttaattgcgc gcttggcgta atcatggtca
     2581 tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga
     2641 agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg
     2701 cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc
     2761 caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac
     2821 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2881 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2941 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3001 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3061 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3121 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3181 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3241 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3301 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3361 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaagg
     3421 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3481 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3541 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3601 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3661 ttcacctaga tcctttttat tcgcggcgtt gtgacaattt accgaacaac tccgcggccg
     3721 ggaagccgat ctcggcttga acgaattgtt aggtggcggt acttgggtcg atatcaaagt
     3781 gcatcacttc ttcccgtatg cccaactttg tatagagagc cactgcggga tcgtcaccgt
     3841 aatctgcttg cacgtagatc acataagcac caagcgcgtt ggcctcatgc ttgaggagat
     3901 tgatgagcgc ggtggcaatg ccctgcctcc ggtgctcgcc ggagactgcg agatcataga
     3961 tatagatctc actacgcggc tgctcaaacc tgggcagaac gtaagccgcg agagcgccaa
     4021 caaccgcttc ttggtcgaag gcagcaagcg cgatgaatgt cttactacgg agcaagttcc
     4081 cgaggtaatc ggagtccggc tgatgttggg agtaggtggc tacgtctccg aactcacgac
     4141 cgaaaagatc aagagcagcc cgcatggatt tgacttggtc agggccgagc ctacatgtgc
     4201 gaatgatgcc catacttgag ccacctaact ttgttttagg gcgactgccc tgctgcgtaa
     4261 catcgttgct gctgcgtaac atcgttgctg ctccataaca tcaaacatcg acccacggcg
     4321 taacgcgctt gctgcttgga tgcccgaggc atagactgta caaaaaaaca gtcataacaa
     4381 gccatgaaaa ccgccactgc gccgttacca ccgctgcgtt cggtcaaggt tctggaccag
     4441 ttgcgtgagc gcatacgcta cttgcattac agtttacgaa ccgaacaggc ttatgtcaac
     4501 tgggttcgtg ccttcatccg tttccacggt gtgcgtcacc cggcaacctt gggcagcagc
     4561 gaagtcgagg catttctgtc ctggctggcg aacgagcgca aggtttcggt ctccacgcat
     4621 cgtcaggcat tggcggcctt gctgttcttc tacggcaagg tgctgtgcac ggatctgccc
     4681 tggcttcagg agatcggaag acctcggccg tcgcggcgct tgccggtggt gctgaccccg
     4741 gatgaagtgg ttcgcatcct cggttttctg gaaggcgagc atcgtttgtt cgcccaggac
     4801 tctagctata gttctagtgg ttggctacga ttattgaagc atttatcagg gttattgtct
     4861 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     4921 atttccccga aaagtgccac ctgacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     4981 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     5041 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     5101 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     5161 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     5221 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     5281 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     5341 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc tt
//
