LOCUS       sequence-374566-        4424 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expression of Luz-eGFP via a T7 promoter in TXTL.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4424)
  AUTHORS   Sato W, Rasmussen M, Deich C, Engelhart AE, Adamala KP
  TITLE     Expanding luciferase reporter systems for cell-free protein 
            expression.
  JOURNAL   Sci Rep. 2022 Jul 7;12(1):11489. doi: 10.1038/s41598-022-15624-6.
  PUBMED    35798760
REFERENCE   2  (bases 1 to 4424)
  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: "Sci Rep."; 
            date: "2022-07-7"; volume: "12(1):11489. doi"; pages: " 
            10.1038/s41598-022-15624-6"
COMMENT     Sequence Label: pTXTL-T7max-LuzGFPHis
FEATURES             Location/Qualifiers
     source          1..4424
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      710..1298
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1199..1218
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1377..1396
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1377..1395
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             1404..1426
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     primer_bind     complement(2306..2327)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2567..2586)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2914..2935
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     CDS             2966..2989
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     rep_origin      3211..3666
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3298..3317)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3508..3529
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3733..3752)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3852..3874
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3912..3930)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3998..4102
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4103..4171,4172..4424,1..539)
                     /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(4321..4340)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
       61 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
      121 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
      181 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
      241 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
      301 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
      361 ttattgctga taaatctgga gccggtgagc gtgggtcgcg cggtatcatt gcagcactgg
      421 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
      481 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
      541 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
      601 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
      661 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
      721 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
      781 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
      841 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
      901 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
      961 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     1021 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     1081 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     1141 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     1201 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     1261 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     1321 tacggttcct ggccttttgc tggccttttg ctcacatggc tcgacagatc taattctaat
     1381 acgactcact atagggaaat aattttgttt aactttaaga aggagatata ccggtatgac
     1441 tagtatgcgt atcaacatct ccctgtcgtc gttattcgaa cggttaagta aactgagctc
     1501 tcgttcaatt gcgattacct gtggagttgt cctcgcctct gctattgctt ttcccatcat
     1561 acgtcgggat tatcagacgt tcctggaggt tggtcctagc tatgccccgc aaaactttcg
     1621 cggctacatt atcgtttgcg tacttagctt gttccgccaa gagcagaaag gcctggcgat
     1681 ctacgaccgg ctcccagaga agcgccgctg gctggcggat ctgccatttc gtgaaggtac
     1741 cagaccgagt attacctcac acattatcca gcgccagcgc actcaacttg tggatcagga
     1801 atttgccaca cgtgaactga ttgacaaggt tattccgagg gtgcaagcac gacataccga
     1861 taaaacattt ttgagcacga gcaagtttga atttcatgcg aaagccattt tcctgttgcc
     1921 atcgattcca atcaacgacc cgctgaacat acctagtcat gatacggtgc gtcgaacgaa
     1981 acgcgaaatt gcccacatgc atgactatca cgattgcacc cttcacctag ctctggcggc
     2041 acaggatgga aaagaagtcc tgaaaaaagg ctggggccag cgtcatccgc tggcggggcc
     2101 gggtgtgcct gggccgccga ctgaatggac tttcttatac gcaccccgta atgaggaaga
     2161 agcgcgcgtg gtagaaatga ttgtggaagc atccatcggc tatatgacca atgatccggc
     2221 aggtaaaatc gtcgagaatg ccaaaggagg tggtggatcg ggcggagggg gatcaatgga
     2281 gcttttcact ggcgttgttc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     2341 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     2401 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     2461 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     2521 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2581 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2641 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2701 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     2761 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2821 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2881 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2941 cgccgggatc acgcgtggag gtggccatca ccatcatcac catcaccacg gataactcga
     3001 gcaaagcccg ccgaaaggcg ggcttttctg tacaacaaca attgcattca ttttatgttt
     3061 caggttcagg gggagatgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     3121 aaaatcgata aggatccggg ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     3181 caacagttgc gcagcctgaa tggcgaatgg acgcgccctg tagcggcgca ttaagcgcgg
     3241 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3301 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3361 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3421 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3481 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3541 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3601 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     3661 caatttcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg
     3721 gtgcactctc agtacaatct gctctgatgc cgcatagtta agccagcccc gacacccgcc
     3781 aacacccgct gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc
     3841 tgtgaccgtc tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc
     3901 gagacgaaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt
     3961 ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt
     4021 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4081 ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4141 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4201 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4261 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4321 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4381 acactattct cagaatgact tggttgagta ctcaccagtc acag
//
