LOCUS       sequence-271800-        3630 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses AausFP2 constitutively in E. coli (most strains).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3630)
  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 3630)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-AausFP2
FEATURES             Location/Qualifiers
     source          1..3630
                     /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(1031..1049)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1045..1092
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1189..1644
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1276..1295)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1486..1507
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1670..1774
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1775..1843,1844..2635)
                     /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(1993..2012)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2806..3394
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3295..3314
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3548..3565
                     /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 gcctgcggag cacttctttt
      301 caaaggaaaa atcccattcg ttgttgaatt agaaggcgat gtaaacggcg aagttttctc
      361 cgtcagaggt aatggttggg gtgatggaag cactggaaaa cttgaaatca aatttgtctg
      421 tacaactggc gaggttcctt tagcttggga atcactcatc tgttccatgg cgtatggtgc
      481 tttagtcttt tgcaaatacc cgagcaacat caacgacttc ttcaagagca caatgccaag
      541 gggatatatt caagaaagaa aaatctcata tgaaaatgac ggaacatttg acgtacgtca
      601 ggaagtcacc tacgaaaacg gagcacttta caaccgggta aaattcaacg gatctggatt
      661 ccgaaaagac ggcaatgttt tggggaaaaa attggatctc acctctattg gaacatgtat
      721 ctacatcatg ggaaatgacg aaggaaccgg cttaaaaggc gtattcaata aggcattcaa
      781 ggttatcggt ggaaaccgtc aaatagccag ccatgtacaa acacaaacac caattggcga
      841 tggattggtt tccattccag actatcatgt tatgcacaat catatcgcat gcagtaaaga
      901 cccgtcagaa acaagagacc acatcattgt aaaagaatcc ttaagagcag tcgattgcaa
      961 cactgaatac atgtgacgtt tgatccggct gctaacaaag cccgaaagga agctgagttg
     1021 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     1081 aggggttttt tgctgaaagg aggaactata tccggatctg gcgtaatagc gaagaggccc
     1141 gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg cgaatgggac gcgccctgta
     1201 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     1261 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     1321 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     1381 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     1441 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     1501 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     1561 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     1621 acaaaatatt aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1681 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1741 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1801 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1861 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1921 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1981 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact
     2041 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     2101 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     2161 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     2221 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     2281 agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg
     2341 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     2401 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     2461 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     2521 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     2581 tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc
     2641 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2701 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2761 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2821 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2881 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2941 accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     3001 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     3061 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcgga
     3121 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     3181 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     3241 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     3301 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     3361 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     3421 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     3481 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     3541 cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct
     3601 ccccgcgcgt tggccgattc attaatgcag
//
