LOCUS       Exported                3599 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Encodes the fluorescent protein amFP486/K68M for constitutive 
            expression in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    pThom77.3
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3599)
  AUTHORS   Molina RS, Tran TM, Campbell RE, Lambert GG, Salih A, Shaner NC, 
            Hughes TE, Drobizhev M
  TITLE     Blue-Shifted Green Fluorescent Protein Homologues Are Brighter than 
            Enhanced Green Fluorescent Protein under Two-Photon Excitation.
  JOURNAL   J Phys Chem Lett. 2017 Jun 15;8(12):2548-2554. doi: 
            10.1021/acs.jpclett.7b00960. Epub 2017 May 25.
  PUBMED    28530831
REFERENCE   2  (bases 1 to 3599)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3599
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     196..215
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     449..466
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     551..570
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        551..569
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             695..712
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             716..748
                     /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             752..775
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(1531..1549)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1545..1592
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1689..2144
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1776..1795)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1986..2007
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2170..2274
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2275..3135
                     /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(2493..2512)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3306..295
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
       61 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
      121 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
      181 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
      241 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
      301 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
      361 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
      421 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
      481 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca ggatctcgat
      541 cccgcgaaat taatacgact cactataggg agaccacaac ggtttccctc tagaatcacc
      601 gagtttattc ttgacacctg atgcgatgaa tgatataata ggaaagtact gttttgttta
      661 actttaagaa ggggatatac atatgcgggg ttctcatcat catcatcatc atggtatggc
      721 tagcatgact ggtggacagc aaatgggtcg ggatctgtac gacgatgacg ataaggatcc
      781 ggccctgagc aacaagttca tcggcgacga catgaagatg acctaccaca tggacggctg
      841 cgtgaacggc cactacttca ccgtgaaggg cgagggcaac ggcaagccct acgagggcac
      901 ccagaccagc accttcaagg tgaccatggc caacggcggc cccctggcct tcagcttcga
      961 catcctgagc accgtgttca tgtacggcaa ccggtgcttc accgcctacc ccaccagcat
     1021 gcccgactac ttcaagcagg ccttccccga cggcatgagc tacgagcgga ccttcaccta
     1081 cgaggacggc ggcgtggcca ccgccagctg ggagatcagc ctgaagggca actgcttcga
     1141 gcacaagagc accttccacg gcgtgaactt ccccgccgac ggccccgtga tggccaagaa
     1201 gaccaccggc tgggacccca gcttcgagaa gatgaccgtg tgcgacggca tcctgaaggg
     1261 cgacgtgacc gccttcctga tgctgcaggg cggcggcaac taccggtgcc agttccacac
     1321 cagctacaag accaagaagc ccgtgaccat gccccccaac cacgtggtgg agcaccggat
     1381 cgcccggacc gacctggaca agggcggcaa cagcgtgcag ctgaccgagc acgccgtggc
     1441 ccacatcacc agcgtggtgc ccttctaaga attcgaagct tgatccggct gctaacaaag
     1501 cccgaaagga agctgagttg gctgctgcca ccgctgagca ataactagca taaccccttg
     1561 gggcctctaa acgggtcttg aggggttttt tgctgaaagg aggaactata tccggatctg
     1621 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     1681 cgaatgggac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     1741 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     1801 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     1861 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
     1921 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     1981 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
     2041 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     2101 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttaggtgg cacttttcgg
     2161 ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     2221 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt
     2281 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     2341 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     2401 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     2461 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     2521 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     2581 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     2641 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     2701 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     2761 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     2821 gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     2881 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     2941 cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt
     3001 atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     3061 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     3121 attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa
     3181 cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa
     3241 atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga
     3301 tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg
     3361 ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact
     3421 ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac
     3481 cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg
     3541 gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttacc
//
