LOCUS       sequence-6714-ex        3080 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 3080)
  AUTHORS   Hartley RW
  TITLE     Directed mutagenesis and barnase-barstar recognition.
  JOURNAL   Biochemistry 1993 Jun 15;32(23):5978-84.
  PUBMED    8507637
REFERENCE   2  (bases 1 to 3080)
  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: 
            "Biochemistry"; date: "1993-06-15"; volume: "32(23)"; pages: 
            "5978-84"
COMMENT     Sequence Label: pMT643
FEATURES             Location/Qualifiers
     source          1..3080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(35..40,41..56,57..63)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    71..87
                     /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     complement(464..480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(464..480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     protein_bind    488..504
                     /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(512..518,519..536,537..542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    557..578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(695..712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(866..1454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(946..965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1625..2416,2417..2485))
                     /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     2248..2267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2486..2590)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2658..2676
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(2714..2736)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2836..2855
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3049..3071
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3063..3080
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3064..3080
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 gaattcgagc tcgagcttac tccccatccc cctgttgaca attaatcatc ggctcgtata
       61 atgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acaggatccg tcataagaaa
      121 ggagccgcac atgaaaaaag cagtcattaa cggggaacaa atcagaagta tcagcgacct
      181 ccaccagaca ttgaaaaagg agcttgccct tccggaatac tacggtgaaa acctggacgc
      241 tttatgggat gcgctgaccg gatgggtgga gtacccgctc gttttggaat ggaggcagtt
      301 tgaacaaagc aagcagctga ctgaaaatgg cgccgagagt gtgcttcagg ttttccgtga
      361 agcgaaagcg gaaggcgcgg acatcaccat catactttct taatacgatc aatgggagat
      421 gaacaatatg gaaacacaaa cccgcaagct tggcgtaatc atggtcatag ctgtttcctg
      481 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
      541 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
      601 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
      661 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
      721 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      781 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      841 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      901 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      961 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     1021 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca atgctcacgc tgtaggtatc
     1081 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     1141 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     1201 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     1261 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     1321 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     1381 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     1441 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     1501 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     1561 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     1621 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1681 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1741 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1801 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1861 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1921 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1981 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     2041 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     2101 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     2161 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     2221 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     2281 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     2341 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     2401 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     2461 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     2521 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     2581 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     2641 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg
     2701 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     2761 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     2821 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     2881 aaataccgca cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc
     2941 tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga
     3001 aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac
     3061 gttgtaaaac gacggccagt
//
