LOCUS       sequence-350353-        3593 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of deGFP via a T7 Promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3593)
  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 3593)
  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-deGFP-8xHis-T500
FEATURES             Location/Qualifiers
     source          1..3593
                     /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)"
     primer_bind     complement(134..155)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(395..414)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     742..763
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             794..817
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     rep_origin      1039..1494
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1126..1145)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1336..1357
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1561..1580)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1680..1702
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(1740..1758)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1826..1930
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1931..1999,2000..2791)
                     /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(2149..2168)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2962..3550
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3451..3470
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccttttgc tcacatggct cgacagatct aattctaata cgactcacta tagggaaata
       61 attttgttta actttaagaa ggagatatac cggtatgact agtatggagc ttttcactgg
      121 cgttgttccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt tcagcgtgtc
      181 cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca tctgcaccac
      241 cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg gcgtgcagtg
      301 cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg ccatgcccga
      361 aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca agacccgcgc
      421 cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg gcatcgactt
      481 caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca gccacaacgt
      541 ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga tccgccacaa
      601 catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc ccatcggcga
      661 cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc tgagcaaaga
      721 ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg ccgggatcac
      781 gcgtggaggt ggccatcacc atcatcacca tcaccacgga taactcgagc aaagcccgcc
      841 gaaaggcggg cttttctgta caacaacaat tgcattcatt ttatgtttca ggttcagggg
      901 gagatgtggg aggtttttta aagcaagtaa aacctctaca aatgtggtaa aatcgataag
      961 gatccgggct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1021 agcctgaatg gcgaatggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1081 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1141 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1201 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1261 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1321 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1381 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1441 tgatttaaca aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttcctgat
     1501 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag
     1561 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     1621 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     1681 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     1741 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     1801 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     1861 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     1921 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     1981 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2041 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2101 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2161 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2221 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2281 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2341 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2401 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2461 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     2521 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     2581 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     2641 gcgtgggtcg cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     2701 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     2761 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     2821 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     2881 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     2941 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3001 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3061 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     3121 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3181 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3241 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3301 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     3361 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3421 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3481 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     3541 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gct
//
