LOCUS       sequence-271810-        3657 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses AausGFP constitutively in E. coli (most strains).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3657)
  AUTHORS   Lambert GG, Depernet H, Gotthard G, Schultz DT, Navizet I, Lambert 
            T, Adams SR, Torreblanca-Zanca A, Chu M, Bindels DS, Levesque V, 
            Nero Moffatt J, Salih A, Royant A, Shaner NC
  TITLE     Aequorea's secrets revealed: New fluorescent proteins with unique 
            properties for bioimaging and biosensing.
  JOURNAL   PLoS Biol. 2020 Nov 2;18(11):e3000936. doi: 
            10.1371/journal.pbio.3000936. eCollection 2020 Nov.
  PUBMED    33137097
REFERENCE   2  (bases 1 to 3657)
  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: "PLoS 
            Biol."; date: "2020-11-2"; volume: "18(11):e3000936. doi"; pages: " 
            10.1371/journal.pbio.3000936. eCollection 2020 Nov"
COMMENT     Sequence Label: pNCST-AausGFP
FEATURES             Location/Qualifiers
     source          1..3657
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     20..39
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        20..38
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             122..144
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             164..181
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             185..217
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             257..277
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     primer_bind     complement(1058..1076)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1072..1119
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1216..1671
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1303..1322)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1513..1534
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1697..1801
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1802..1870,1871..2662)
                     /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(2020..2039)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2833..3421
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3322..3341
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3575..3592
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 gatctcgatc ccgcgaaatt aatacgactc actataggga gaccacaacg gtttccctct
       61 agaatcaccg agtttattct tgacacctga tgcgatgaat gatataatag gaaagtactg
      121 ttttgtttaa ctttaagaag gagatataca tatgcggggt tctcatcatc atcatcatca
      181 tggtatggct agcatgactg gtggacagca aatgggtcgg gatctgtacg acgatgacga
      241 taacgatccg ctcgaggaaa acctgtactt ccagggtatg aagatttatt gcgaaggaga
      301 aaagctattc aaggggattg ttccaatcca aattgaattg aatggtgatg tcaatggtca
      361 caagttttct gtccatggtg aagggcaagg agaagcagag atgggtaggc ttaccctcag
      421 attcgtttgt actactggaa agatgcctgt accatggcca acacttgtca ctacattctc
      481 ttatggtgta caatgttttt caaggtaccc agagcacatg aaaatgcacg acttcttcaa
      541 gagtgccatg cctgaaggat atacacaaga gcgtaccatc tttttcgata aagatggtac
      601 ctacaagacg cgtgctgagg tcaagttcga aggtgataca cttgttaaca ggattgagtt
      661 gacaggtact ggattcaagg aagatggtaa tattcttggg cataagatgg aatacaatta
      721 caattcacat aatgtttaca ttatgtcgga taaagcaaag aacggaatca aagtcaactt
      781 caaaatcagg cataacattg aagatggtac agtacaattg gctgatcatt accaagcaaa
      841 ttatccactc ggtaaaggcc ctgtccttct accagaagat cactacttgt ctactcaatc
      901 tgcccttacc aaagatcccc acgaaacaag agatcacatg ttactattgg agtttgttac
      961 tgctgctggt atcacacatg gaatggatga gctatacaag tgacgtttga tccggctgct
     1021 aacaaagccc gaaaggaagc tgagttggct gctgccaccg ctgagcaata actagcataa
     1081 ccccttgggg cctctaaacg ggtcttgagg ggttttttgc tgaaaggagg aactatatcc
     1141 ggatctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgcagcc
     1201 tgaatggcga atgggacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta
     1261 cgcgcagcgt gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc
     1321 cttcctttct cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt
     1381 tagggttccg atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg
     1441 gttcacgtag tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca
     1501 cgttctttaa tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct
     1561 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     1621 tttaacaaaa atttaacgcg aattttaaca aaatattaac gcttacaatt taggtggcac
     1681 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     1741 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     1801 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     1861 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     1921 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     1981 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     2041 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     2101 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     2161 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     2221 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     2281 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     2341 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     2401 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     2461 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     2521 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     2581 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     2641 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     2701 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     2761 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     2821 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     2881 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     2941 ggtaactggc ttcagcagag cgcagatacc aaatactgtt cttctagtgt agccgtagtt
     3001 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     3061 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     3121 gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcacac agcccagctt
     3181 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     3241 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     3301 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     3361 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     3421 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     3481 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     3541 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     3601 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcag
//
