LOCUS       sequence-452398-        3152 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pT7-driven expression of alpha-hemolysin, a pore-forming protein in 
            lipid bilayers..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3152)
  AUTHORS   Murray, RM
  TITLE     Membrane proteins
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 3152)
  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: 
            "Unpublished"
COMMENT     Sequence Label: PT7-alphaHL
FEATURES             Location/Qualifiers
     source          1..3152
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     41..59
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(63..106)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    109..130
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     primer_bind     150..169
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        150..168
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             247..255
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     terminator      1229..1276
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    1289..1309
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1310..1367
                     /label=his operon terminator
                     /note="This putative transcriptin terminator from the E. 
                     coli his operon has a 2-bp deletion introduced during 
                     synthesis. Its efficiency has not been determined."
     primer_bind     complement(1391..1408)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     complement(1616..1635)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1627..2442
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1700..1719)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2564..3152)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2644..2663)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 gaagatcctt tgatcttttc tacggggtct gacgcgcata aataggcgta tcacgaggca
       61 gaatttcaga taaaaaaaat ccttagcttt cgctaaggat gatttctgga attcgcggcc
      121 gcttctagag actagtggaa gacatggagt aatacgactc actataggga gatactgggc
      181 ccaagttcac ttaaaaagga gatcaacaat gaaagcaatt ttcgtactga aacatcttaa
      241 tcatgctaag gaggttttct aatgatgaaa acacgtatag tcagctcagt aacaacaaca
      301 ctattgctag gttccatatt aatgaatcct gtcgctaatg ccgcagattc tgatattaat
      361 attaaaaccg gtactacaga tattggaagc aatactacag taaaaacagg tgatttagtc
      421 acttatgata aagaaaatgg catgcacaaa aaagtatttt atagttttat cgatgataaa
      481 aatcataata aaaaactgct agttattaga acgaaaggta ccattgctgg tcaatataga
      541 gtttatagcg aagaaggtgc taacaaaagt ggtttagcct ggccttcagc ctttaaggta
      601 cagttgcaac tacctgataa tgaagtagct caaatatctg attactatcc aagaaattcg
      661 attgatacaa aagagtatat gagtacttta acttatggat tcaacggtaa tgttactggt
      721 gatgatacag gaaaaattgg cggccttatt ggtgcaaatg tttcgattgg tcatacactg
      781 aaatatgttc aacctgattt caaaacaatt ttagagagcc caactgataa aaaagtaggc
      841 tggaaagtga tatttaacaa tatggtgaat caaaattggg gaccatatga tagagattct
      901 tggaacccgg tatatggcaa tcaacttttc atgaaaacta gaaatggctc tatgaaagca
      961 gcagataact tccttgatcc taacaaagca agttctctat tatcttcagg gttttcacca
     1021 gacttcgcta cagttattac tatggataga aaagcatcca aacaacaaac aaatatagat
     1081 gtaatatacg aacgagttcg tgatgactac caattgcact ggacttcaac aaattggaaa
     1141 ggtaccaata ctaaagataa atggatagat cgttcttcag aaagatataa aatcgattgg
     1201 gaaaaagaag aaatgacaaa ttaaaggtct agcataaccc cttggggcct ctaaacgggt
     1261 cttgaggggt tttttggctt atgtcttcta ctagtagcgg ccgctgcagt ccggcaaaaa
     1321 agggcaaggt gtcaccaccc tgcccttttt ctttaaaacc gaaaagatta cttcgcgtta
     1381 tgcaggcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     1441 cagctcactc aaaggcggta agtcgcgtgt tatacgcccg ttatccatgg gtatggacag
     1501 ttttcccttt gatatgtaac gcacgttgtg tctcaaaatc tctgatgtta cattgcacaa
     1561 gataaaaata tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     1621 ggtgttatga gccatattca acgggaaacg tcttgctccc gtccgcgctt aaactccaac
     1681 atggacgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     1741 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     1801 ggtagcgttg ccaatgatgt tacagatgag atggtccgtc tcaactggct gacggagttt
     1861 atgcctctcc cgaccatcaa gcattttatc cgtactcctg atgatgcgtg gttactcacc
     1921 accgcgattc ctgggaaaac agccttccag gtattagaag aatatcctga ttcaggtgaa
     1981 aatattgttg atgcgctggc cgtgttcctg cgccggttac attcgattcc tgtttgtaat
     2041 tgtcctttta acagcgatcg tgtatttcgt cttgctcagg cgcaatcacg catgaataac
     2101 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2161 tggaaagaaa tgcacaagct cttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2221 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2281 cgggtcggaa tcgcagaccg ttaccaggac cttgccattc tttggaactg cctcggtgag
     2341 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2401 aataaattgc agtttcattt gatgctcgat gagtttttct aataatacta gcagaaatca
     2461 tccttagcga aagctaagga ttttttttat ctgattaccg cctttgagtg agcgtcgacc
     2521 tagtcctagc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     2581 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     2641 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2701 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2761 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2821 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2881 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2941 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     3001 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     3061 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     3121 agcagattac gcgcagaaaa aaaggatctc aa
//
