LOCUS       sequence-7375-ex        4980 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 4980)
  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 4980)
  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: pB1H2
FEATURES             Location/Qualifiers
     source          1..4980
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(89..94,95..112,113..119)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    127..143
                     /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     132..154
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     151..167
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     151..167
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             966..989
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     rep_origin      2041..2496
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2128..2147)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2338..2359
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2522..2626
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2627..2695,2696..3487)
                     /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     complement(2845..2864)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3658..4246
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4147..4166
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4400..4417
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4432..4572
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4498..4517)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4658..4680
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4674..4865)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 gaattgcaaa aaaatattga caacataaaa aactttgtgt tatacttgta acgctacatg
       61 gagattaact caatctagct agagaggctt tacactttat gcttccggct cgtataatgt
      121 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacggat
      181 tcactggaac tctaaaccaa agagaggaca caatgcaggg ttctgtgaca gagtttctaa
      241 aaccgcgcct ggttgatatc gagcaagtga gttcgacgca cgccaaggtg acccttgagc
      301 ctttagagcg tggctttggc catactctgg gtaacgcact gcgccgtatt ctgctctcat
      361 cgatgccggg ttgcgcggtg accgaggttg agattgatgg tgtactacat gagtacagca
      421 ccaaagaagg cgttcaggaa gatatcctgg aaatcctgct caacctgaaa gggctggcgg
      481 tgagagttca gggcaaagat gaagttattc ttaccttgaa taaatctggc attggccctg
      541 tgactgcagc cgatatcacc cacgacggtg atgtcgaaat cgtcaagccg cagcacgtga
      601 tctgccacct gaccgatgag aacgcgtcta ttagcatgcg tatcaaagtt cagcgcggtc
      661 gtggttatgt gccggcttct acccgaattc attcggaaga agatgagcgc ccaatcggcc
      721 gtctgctggt cgacgcatgc tacagccctg tggagcgtat tgcctacaat gttgaagcag
      781 cgcgtgtaga acagcgtacc gacctggaca agctggtcat cgaaatggaa accaacggca
      841 caatcgatcc tgaagaggcg attcgtcgtg cggcaaccat tctggctgaa caactggaag
      901 cgttcgttga cttacgtgat gtacgtcagc ctgaagtgaa agaagagaaa ccagaggcgg
      961 ccgcggacta caaggatgac gacgacaagt tccggaccgg ttccaagaca cccccccatg
     1021 gtaccaagcc ctacgtaaag atcaccgaac aaccggcagg aaaggcgcta aggtttcgct
     1081 acgagtgcga gggacgctcg gcgggatcca ttccgggcgt gaactctacg ccggagaaca
     1141 agacctatcc gacaatcgaa attgtgggct acaagggacg cgccgtagtt gttgtctcct
     1201 gcgtcacaaa ggatacgcca tatcgtcctc atccccacaa tttagttggc aaggagggct
     1261 gcaagaaggg cgtctgtaca ctggagatca acagtgagac aatgcgagcg gtgttcagta
     1321 acttgggtat ccagtgtgtc aaaaagaagg acattgaggc ggcgctcaag gcgcgcgagg
     1381 agatccgtgt ggatccgttt aagactggct tttcgcatcg tttccagccc tcgagcatag
     1441 atctgaattc ggtgcgattg tgctttcaag tattcatgga gagcgagcag aagggtcgat
     1501 tcacctcgcc actgccgccg gtagtttcgg agcccatctt cgataagaag gccatgtccg
     1561 acctggtcat ctgccggctg tgtagctgct cggccaccgt tttcggcaac acccagatca
     1621 tcctgctctg cgagaaggtg gccaaggagg acatctccgt gcgattcttt gaggagaaga
     1681 atggccagag tgtttgggag gcctttggtg acttccagca cacggatgtc cacaagcaga
     1741 ctgccattac ctttaagacg ccgcgctatc ataccctgga catcacagag cccgccaagg
     1801 tttttattca actgcgacgt ccctcggatg gagttaccag cgaggccctg cccttcgagt
     1861 acgtgccaat ggactcagat ccagcgcact tgaggcggaa acgtcagaag taacctagga
     1921 tcctctagag actagaaaaa ggccgacaag tcccgctccg ctgaagatcc tggcgtaata
     1981 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggg
     2041 acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg
     2101 ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca
     2161 cgttcgccgg ctttccccgt caagctctaa atcgggggct ccctttaggg ttccgattta
     2221 gtgctttacg gcacctcgac cccaaaaaac ttgattaggg tgatggttca cgtagtgggc
     2281 catcgccctg atagacggtt tttcgccctt tgacgttgga gtccacgttc tttaatagtg
     2341 gactcttgtt ccaaactgga acaacactca accctatctc ggtctattct tttgatttat
     2401 aagggatttt gccgatttcg gcctattggt taaaaaatga gctgatttaa caaaaattta
     2461 acgcgaattt taacaaaata ttaacgctta caatttaggt ggcacttttc ggggaaatgt
     2521 gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag
     2581 acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca
     2641 tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc
     2701 agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat
     2761 cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc
     2821 aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg
     2881 gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc
     2941 agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat
     3001 aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga
     3061 gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc
     3121 ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc
     3181 aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt
     3241 aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc
     3301 tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc
     3361 agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca
     3421 ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca
     3481 ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa aacttcattt
     3541 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     3601 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3661 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3721 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3781 cagagcgcag ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa
     3841 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3901 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3961 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     4021 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     4081 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     4141 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     4201 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     4261 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     4321 atcccctgat tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg
     4381 cagccgaacg accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg
     4441 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
     4501 aatctgctct gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg
     4561 gtcatggctg cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg
     4621 ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg
     4681 ttttcaccgt catcaccgaa acgcgcgagg cagctgcggt aaagctcatc agcgtggtcg
     4741 tgaagcgatt cacagatgtc tgcctgttca tccgcgtcca gctcgttgag tttctccaga
     4801 agcgttaatg tctggcttct gataaagcgg gccatgttaa gggcggtttt ttcctgtttg
     4861 gtcactgatg cctccgtgta agggggattt ctgttcatgg gggtaatgat accgatgaaa
     4921 cgagagagga tgctcacgat acgggttact gatgatgaac atgcccggtt actggaacgg
//
