LOCUS       sequence-350323-        2930 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Empty backbone. T7Max promoter with N-His Tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2930)
  AUTHORS   Deich C, Cash B, Sato W, Sharon J, Aufdembrink L, Gaut NJ, Heili J, 
            Stokes K, Engelhart AE, Adamala KP
  TITLE     T7Max transcription system.
  JOURNAL   J Biol Eng. 2023 Jan 23;17(1):4. doi: 10.1186/s13036-023-00323-1.
  PUBMED    36691081
REFERENCE   2  (bases 1 to 2930)
  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: "J Biol 
            Eng."; date: "2023-01-23"; volume: "17(1):4. doi"; pages: " 
            10.1186/s13036-023-00323-1"
COMMENT     Sequence Label: pCI-T7Max-UTR1-NTerminus8xHis-T500
FEATURES             Location/Qualifiers
     source          1..2930
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     36..55
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        36..54
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             63..85
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             101..124
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     rep_origin      376..831
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(463..482)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     673..694
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(898..917)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1017..1039
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1077..1095)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1163..1267
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1268..1336,1337..2128)
                     /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(1486..1505)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2299..2887
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2788..2807
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatggct cgacagatct aattctaata cgactcacta tagggaaata
       61 attttgttta actttaagaa ggagatatac cggtatgggc catcaccatc atcaccatca
      121 ccacggaggt ggcactagtg aattccgtac gacgcgttaa ctcgagcaaa gcccgccgaa
      181 aggcgggctt ttctgtacaa caacaattgc attcatttta tgtttcaggt tcagggggag
      241 atgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtaaaat cgataaggat
      301 ccgggctggc gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc
      361 ctgaatggcg aatggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
      421 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
      481 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
      541 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
      601 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
      661 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
      721 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
      781 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt tcctgatgcg
      841 gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac
      901 aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc
      961 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     1021 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct
     1081 cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg
     1141 tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc
     1201 aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag
     1261 gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg
     1321 ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt
     1381 gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt
     1441 tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt
     1501 attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa
     1561 tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag
     1621 agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac
     1681 aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac
     1741 tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac
     1801 cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac
     1861 tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact
     1921 tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg
     1981 tgggtcgcgc ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt
     2041 tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat
     2101 aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta
     2161 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     2221 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     2281 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     2341 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     2401 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     2461 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     2521 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     2581 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     2641 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2701 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2761 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2821 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2881 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
//
