LOCUS       sequence-333656-        2509 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reverse Ptac promoter part (H).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2509)
  AUTHORS   Greco FV, Pandi A, Erb TJ, Grierson CS, Gorochowski TE
  TITLE     Harnessing the central dogma for stringent multi-level control of 
            gene expression.
  JOURNAL   Nat Commun. 2021 Mar 19;12(1):1738. doi: 10.1038/s41467-021-21995-7.
  PUBMED    33741937
REFERENCE   2  (bases 1 to 2509)
  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 
            Commun."; date: "2021-03-19"; volume: "12(1):1738. doi"; pages: " 
            10.1038/s41467-021-21995-7"
COMMENT     Sequence Label: pH-01
FEATURES             Location/Qualifiers
     source          1..2509
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      580..1168
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1069..1088
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1261..1278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(1486..1491,1492..1507,1508..1514)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    1520..1539
                     /label=lac operator (symmetric)
                     /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). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     primer_bind     complement(1591..1610)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1592..1610)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1621..1638)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1621..1637)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1630..1652)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        1953..2057
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2058..2126,2127..2509,1..409)
                     /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(2276..2295)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
       61 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
      121 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
      181 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
      241 taaatctgga gccggtgagc gtggttctcg cggtatcatt gcagcactgg ggccagatgg
      301 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
      361 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
      421 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
      481 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
      541 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
      601 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
      661 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
      721 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
      781 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
      841 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
      901 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
      961 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     1021 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     1081 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     1141 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     1201 ggccttttgc tggccttttg ctcattaggc accccaggct ttacccgaac gaccgagcgc
     1261 agcgagtcag tgagcgagga agcggagagc gcccaatacg caaggaaaca gctatgacca
     1321 tgttaatgca gctggcacga caggtttccc gactggaaag cgggcagtga gcggaaggcc
     1381 catgaggccc agagggtacc atgaccatta acgtccccat ccaactcaac caagatgggt
     1441 acaacgcctg taaacagctc ctctccttta ctggtctcag gcatgttgac aattaatcat
     1501 cggctcgtat aatgtgtgga attgtgagcg ctcacaatta gtatgagacc gagctcgttg
     1561 atgcggcctt gacggccttc cgccaattcg ccctatagtg agtcgtatta cgtcgcgctc
     1621 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     1681 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgaaac
     1741 gcccttccca acagttgcgc agcctgaatg gcgaatggga gcgccctgta gcggccactc
     1801 aaccctatct cggtctattc ttttgattta taagggattt tgccgatttc ggcctattgg
     1861 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     1921 acaatttagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     1981 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2041 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     2101 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     2161 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     2221 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     2281 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     2341 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     2401 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     2461 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgca
//
