LOCUS       sequence-403118-        4799 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid for the expression of the basic region and leucine zipper 
            domain of the Saccharomyces cerevisiae transcription factor GCN4 in 
            bacterial cells.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4799)
  AUTHORS   Weiss MA, Ellenberger T, Wobbe CR, Lee JP, Harrison SC, Struhl K
  TITLE     Folding transition in the DNA-binding domain of GCN4 on specific 
            binding to DNA.
  JOURNAL   Nature. 1990 Oct 11;347(6293):575-8. doi: 10.1038/347575a0.
  PUBMED    2145515
REFERENCE   2  (bases 1 to 4799)
  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"; journalName: 
            "Nature"; date: "1990-10-11- 11"; volume: "347"; issue: "6293"; 
            pages: "575-8"
COMMENT     Sequence Label: pGCNK58
FEATURES             Location/Qualifiers
     source          1..4799
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1405..1424
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1508..1527
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1508..1526
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1558..1580
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1690..1746
                     /codon_start=1
                     /product="GCN4 peptide optimized to improve solubility 
                     while preserving binding to an scFv-GFP fusion protein 
                     (Tanenbaum et al., 2014)"
                     /label=GCN4_v4
                     /translation="EELLSKNYHLENEVARLKK"
     primer_bind     complement(1836..1854)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1850..1897
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(2263..2268,2269..2285,2286..2291))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(2318..2336)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2404..2508
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2509..2577,2578..3369)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2727..2746)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3540..4128
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4029..4048
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4282..4299
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4314..4453
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4379..4398)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4539..4561
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4555..4746)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
ORIGIN
        1 aacgagagag gatgctcacg atacgggtta ctgatgatga acatgcccgg ttactggaac
       61 gttgtgaggg taaacaactg gcggtatgga tgcggcggga ccagagaaaa atcactcagg
      121 gtcaatgcca gcgcttcgtt aatacagatg taggtgttcc acagggtagc cagcagcatc
      181 ctgcgatgca gatccggaac ataatggtgc agggcgctga cttccgcgtt tccagacttt
      241 acgaaacacg gaaaccgaag accattcatg ttgttgctca ggtcgcagac gttttgcagc
      301 agcagtcgct tcacgttcgc tcgcgtatcg gtgattcatt ctgctaacca gtaaggcaac
      361 cccgccagcc tagccgggtc ctcaacgaca ggagcacgat catgcgcacc cgtggccagg
      421 acccaacgct gcccgagatg cgccgcgtgc ggctgctgga gatggcggac gcgatggata
      481 tgttctgcca agggttggtt tgcgcattca cagttctccg caagaattga ttggctccaa
      541 ttcttggagt ggtgaatccg ttagcgaggt gccgccggct tccattcagg tcgaggtggc
      601 ccggctccat gcaccgcgac gcaacgcggg gaggcagaca aggtataggg cggcgcctac
      661 aatccatgcc aacccgttcc atgtgctcgc cgaggcggca taaatcgccg tgacgatcag
      721 cggtccaatg atcgaagtta ggctggtaag agccgcgagc gatccttgaa gctgtccctg
      781 atggtcgtca tctacctgcc tggacagcat ggcctgcaac gcgggcatcc cgatgccgcc
      841 ggaagcgaga agaatcataa tggggaaggc catccagcct cgcgtcgcga acgccagcaa
      901 gacgtagccc agcgcgtcgg ccgccatgcc ggcgataatg gcctgcttct cgccgaaacg
      961 tttggtggcg ggaccagtga cgaaggcttg agcgagggcg tgcaagattc cgaataccgc
     1021 aagcgacagg ccgatcatcg tcgcgctcca gcgaaagcgg tcctcgccga aaatgaccca
     1081 gagcgctgcc ggcacctgtc ctacgagttg catgataaag aagacagtca taagtgcggc
     1141 gacgatagtc atgccccgcg cccaccggaa ggagctgact gggttgaagg ctctcaaggg
     1201 catcggtcga cgctctccct tatgcgactc ctgcattagg aagcagccca gtagtaggtt
     1261 gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg cgcccaacag
     1321 tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca tgagcccgaa
     1381 gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag caaccgcacc
     1441 tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga tctcgatccc
     1501 gcgaaattaa tacgactcac tatagggaga ccacaacggt ttccctctag aaataatttt
     1561 gtttaacttt aagaaggaga tatacatatg aaagatccgg cggcgctgaa acgcgcgcgt
     1621 aacacagaag ctgcgcgtcg tagccgtgct cgtaaactgc agcgtatgaa gcagctcgag
     1681 gataaagtgg aagaactctt aagcaaaaac taccacctgg aaaacgaagt ggcacgtctg
     1741 aaaaagcttg tgggtgagcg ttaatgagct ctcgagtcta gaatcggatc cggctgctaa
     1801 caaagcccga aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc
     1861 ccttggggcc tctaaacggg tcttgagggg ttttttgctg aaaggaggaa ctatatccgg
     1921 atatccacag gacgggtgtg gtcgccatga tcgcgtagtc gatagtggct ccaagtagcg
     1981 aagcgagcag gactgggcgg cggccaaagc ggtcggacag tgctccgaga acgggtgcgc
     2041 atagaaattg catcaacgca tatagcgcta gcagcacgcc atagtgactg gcgatgctgt
     2101 cggaatggac gatatcccgc aagaggcccg gcagtaccgg cataaccaag cctatgccta
     2161 cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat ttcatacacg
     2221 gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc ttatcgatga
     2281 taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg cctattttta
     2341 taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat
     2401 gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg
     2461 agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa
     2521 catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac
     2581 ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac
     2641 atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt
     2701 ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg tgttgacgcc
     2761 gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca
     2821 ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc
     2881 ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag
     2941 gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa
     3001 ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgcagcaatg
     3061 gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa
     3121 ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg
     3181 gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt
     3241 gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt
     3301 caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag
     3361 cattggtaac tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat
     3421 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     3481 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     3541 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     3601 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     3661 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     3721 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     3781 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     3841 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     3901 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     3961 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     4021 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     4081 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     4141 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     4201 ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc
     4261 cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg
     4321 cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc actctcagta
     4381 caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc tacgtgactg
     4441 ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     4501 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag
     4561 gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat cagcgtggtc
     4621 gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga gtttctccag
     4681 aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt tttcctgttt
     4741 ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga taccgatga
//
