LOCUS       sequence-51172-e        2863 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2863)
  AUTHORS   MacBeath G, Kast P
  TITLE     UGA read-through artifacts--when popular gene expression systems 
            need a pATCH.
  JOURNAL   Biotechniques. 1998 May;24(5):789-94.
  PUBMED    9591128
REFERENCE   2  (bases 1 to 2863)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Biotechniques."; date: "1998-05"; volume: "24(5)"; pages: "789-94"
COMMENT     Sequence Label: pATCH1
FEATURES             Location/Qualifiers
     source          1..2863
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             22..39
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(108..126)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      122..169
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      206..661
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(293..312)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     503..524
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        687..791
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(792..860,861..1652)
                     /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(1010..1029)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1823..2411
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2312..2331
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2565..2582
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2699..2720
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2735..2740,2741..2758,2759..2765)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2773..2789
                     /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     2778..2800
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2797..2813
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2797..2813
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        2834..2852
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ggtaccgggc cccccctcga gcaccaccac caccaccact aataatgact agtcagctga
       61 tccggctgct aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata
      121 actagcataa ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg
      181 aactatatcc ggattggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt
      241 gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc
      301 gctttcttcc cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg
      361 gggctccctt tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat
      421 tagggtgatg gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg
      481 ttggagtcca cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct
      541 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
      601 aatgagctga tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt
      661 taggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
      721 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
      781 aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat
      841 tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc
      901 agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga
      961 gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg
     1021 cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc
     1081 agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag
     1141 taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc
     1201 tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg
     1261 taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg
     1321 acaccacgat gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac
     1381 ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac
     1441 cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg
     1501 agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg
     1561 tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg
     1621 agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac
     1681 tttagattga tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg
     1741 ataatctcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     1801 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     1861 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     1921 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt
     1981 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     2041 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     2101 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     2161 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     2221 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     2281 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     2341 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     2401 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     2461 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     2521 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     2581 aggaagcgga agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt
     2641 aatgcagctg gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta
     2701 atgtgagtta gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta
     2761 tgttgtgtgg aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt
     2821 acgccaagct cgaaattaac cctcactaaa gggaacaaaa gct
//
