LOCUS       sequence-7373-ex        4485 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 4485)
  AUTHORS   Meng X, Brodsky MH, Wolfe SA
  TITLE     A bacterial one-hybrid system for determining the DNA-binding 
            specificity of transcription factors.
  JOURNAL   Nat Biotechnol. 2005 Aug . 23(8):988-94.
  PUBMED    16041365
REFERENCE   2  (bases 1 to 4485)
  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: "Nat 
            Biotechnol. 2005 Aug . 23(8):988-94."
COMMENT     Sequence Label: pB1H1
FEATURES             Location/Qualifiers
     source          1..4485
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     134..153
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(220..322)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     primer_bind     complement(714..731)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(848..1393)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(928..947)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1572..1590
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        join(1842..1847,1848..1865,1866..1872)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    1880..1896
                     /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     1885..1907
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1904..1920
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1904..1920
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             2719..2742
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             complement(join(4045..4485,1..219))
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
ORIGIN
        1 gaattccgga tgagcattca tcaggcgggc aagaatgtga ataaaggccg gataaaactt
       61 gtgcttattt ttctttacgg tctttaaaaa ggccgtaata tccagctgaa cggtctggtt
      121 ataggtacat tgagcaactg actgaaatgc ctcaaaatgt tctttacgat gccattggga
      181 tatatcaacg gtggtatatc cagtgatttt tttctccatt ttagcttcct tagctcctga
      241 aaatctcgat aactcaaaaa atacgcccgg tagtgatctt atttcattat ggtgaaagtt
      301 ggaacctctt acgtgccgat caacgtctca ttttcgccaa aagttggccc agggcttccc
      361 ggtatcaaca gggacaccag gatttattta ttctgcgaag tgatcttccg tcacaggtat
      421 ttattcggcg caaagtgcgt cgggtgatgc tgccaactta ctgatttagt gtatgatggt
      481 gtttttgagg tgctccagtg gcttctgttt ctatcagctg tccctcctgt tcagctactg
      541 acggggtggt gcgtaacggc aaaagcaccg ccggacatca gcgctagcgg agtgtatact
      601 ggcttactat gttggcactg atgagggtgt cagtgaagtg cttcatgtgg caggagaaaa
      661 aaggctgcac cggtgcgtca gcagaatatg tgatacagga tatattccgc ttcctcgctc
      721 actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta cgaacggggc
      781 ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg gccgcggcaa
      841 agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga cgctcaaatc
      901 agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct ggcggctccc
      961 tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg ttatggccgc
     1021 gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc caagctggac
     1081 tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa ctatcgtctt
     1141 gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg taattgattt
     1201 agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga caagttttgg
     1261 tgactgcgct cctccaagcc agttacctcg gttcaaagag ttggtagctc agagaacctt
     1321 cgaaaaaccg ccctgcaagg cggttttttc gttttcagag caagagatta cgcgcagacc
     1381 aaaacgatct caagaagatc atcttattaa tcagataaaa tatttctaga tttcagtgca
     1441 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgtaattctc
     1501 atgtttgaca gcttatcatc gataagctgg acgtctaaga aaccattatt atcatgacat
     1561 taacctataa aaataggcgt atcacgaggc cctttggata accagaagca ataaaaaatc
     1621 aaatcggatt tcactatata atctcacttt atctaagatg aatccgatgg aagcatcctg
     1681 ttttctctca atttttttat ctaaaaccca gcgttcgatg cttctttgag cgaacgatca
     1741 aaaataagtg ccttcccatc aaaaaaatat tgacaacata aaaaactttg tgttatactt
     1801 gtaacgctac atggagatta actcaatcta gctagagagg ctttacactt tatgcttccg
     1861 gctcgtataa tgtgtggaat tgtgagcgga taacaatttc acacaggaaa cagctatgac
     1921 catgattacg gattcactgg aactctagac caaagagagg acacaatgca gggttctgtg
     1981 acagagtttc taaaaccgcg cctggttgat atcgagcaag tgagttcgac gcacgccaag
     2041 gtgacccttg agcctttaga gcgtggcttt ggccatactc tgggtaacgc actgcgccgt
     2101 attctgctct catcgatgcc gggttgcgcg gtgaccgagg ttgagattga tggtgtacta
     2161 catgagtaca gcaccaaaga aggcgttcag gaagatatcc tggaaatcct gctcaacctg
     2221 aaagggctgg cggtgagagt tcagggcaaa gatgaagtta ttcttacctt gaataaatct
     2281 ggcattggcc ctgtgactgc agccgatatc acccacgacg gtgatgtcga aatcgtcaag
     2341 ccgcagcacg tgatctgcca cctgaccgat gagaacgcgt ctattagcat gcgtatcaaa
     2401 gttcagcgcg gtcgtggtta tgtgccggct tctacccgaa ttcattcgga agaagatgag
     2461 cgcccaatcg gccgtctgct ggtcgacgca tgctacagcc ctgtggagcg tattgcctac
     2521 aatgttgaag cagcgcgtgt agaacagcgt accgacctgg acaagctggt catcgaaatg
     2581 gaaaccaacg gcacaatcga tcctgaagag gcgattcgtc gtgcggcaac cattctggct
     2641 gaacaactgg aagctttcgt tgacttacgt gatgtacgtc agcctgaagt gaaagaagag
     2701 aaaccagagg cggccgcgga ctacaaggat gacgacgaca agttccggac cggttccaag
     2761 acaccccccc atggtaccaa gccctacgta aagatcaccg aacaaccggc aggaaaggcg
     2821 ctaaggtttc gctacgagtg cgagggacgc tcggcgggat ccattccggg cgtgaactct
     2881 acgccggaga acaagaccta tccgacaatc gaaattgtgg gctacaaggg acgcgccgta
     2941 gttgttgtct cctgcgtcac aaaggatacg ccatatcgtc ctcatcccca caatttagtt
     3001 ggcaaggagg gctgcaagaa gggcgtctgt acactggaga tcaacagtga gacaatgcga
     3061 gcggtgttca gtaacttggg tatccagtgt gtcaaaaaga aggacattga ggcggcgctc
     3121 aaggcgcgcg aggagatccg tgtggatccg tttaagactg gcttttcgca tcgtttccag
     3181 ccctcgagca tagatctgaa ttcggtgcga ttgtgctttc aagtattcat ggagagcgag
     3241 cagaagggtc gattcacctc gccactgccg ccggtagttt cggagcccat cttcgataag
     3301 aaggccatgt ccgacctggt catctgccgg ctgtgtagct gctcggccac cgttttcggc
     3361 aacacccaga tcatcctgct ctgcgagaag gtggccaagg aggacatctc cgtgcgattc
     3421 tttgaggaga agaatggcca gagtgtttgg gaggcctttg gtgacttcca gcacacggat
     3481 gtccacaagc agactgccat tacctttaag acgccgcgct atcataccct ggacatcaca
     3541 gagcccgcca aggtttttat tcaactgcga cgtccctcgg atggagttac cagcgaggcc
     3601 ctgcccttcg agtacgtgcc aatggactca gatccagcgc acttgaggcg gaaacgtcag
     3661 aagtaaccta ggatcctcta ggatctgcat cgcaggatgc tgctggctac cctgtggaac
     3721 acctacatct gtattaacga agcgctaacc gtttttatca ggctctggga ggcagaataa
     3781 atgatcatat cgtcaattat tacctccacg gggagagcct gagcaaactg gcctcaggca
     3841 tttgagaagc acacggtcac actgcttccg gtagtcaata aaccggtaaa ccagcaatag
     3901 acataagcgg ctatttaacg accctgccct gaaccgacga ccgggtcgaa tttgctttcg
     3961 aatttctgcc attcatccgc ttattatcac ttattcaggc gtagcaccag gcgtttaagg
     4021 gcaccaataa ctgccttaaa aaaattacgc cccgccctgc cactcatcgc agtactgttg
     4081 taattcatta agcattctgc cgacatggaa gccatcacag acggcatgat gaacctgaat
     4141 cgccagcggc atcagcacct tgtcgccttg cgtataatat ttgcccatgg tgaaaacggg
     4201 ggcgaagaag ttgtccatat tggccacgtt taaatcaaaa ctggtgaaac tcacccaggg
     4261 attggctgag acgaaaaaca tattctcaat aaacccttta gggaaatagg ccaggttttc
     4321 accgtaacac gccacatctt gcgaatatat gtgtagaaac tgccggaaat cgtcgtggta
     4381 ttcactccag agcgatgaaa acgtttcagt ttgctcatgg aaaacggtgt aacaagggtg
     4441 aacactatcc catatcacca gctcaccgtc tttcattgcc atacg
//
