LOCUS       Exported                3533 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Encodes the fluorescent protein amFP486 for constitutive expression 
            in E. coli..
ACCESSION   .
VERSION     .
KEYWORDS    pThom77.2
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3533)
  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 3533)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3533
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     complement(63..81)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      77..124
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      221..676
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(308..327)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     518..539
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        702..806
                     /gene="bla"
                     /label=AmpR promoter
     CDS             807..1601
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(861..880)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     1471..1490
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     rep_origin      1772..2360
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2261..2280
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2514..2531
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     2616..2635
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2616..2634
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             2760..2777
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2781..2813
                     /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             2817..2840
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
ORIGIN
        1 gaattcgaag cttgatccgg ctgctaacaa agcccgaaag gaagctgagt tggctgctgc
       61 caccgctgag caataactag cataacccct tggggcctct aaacgggtct tgaggggttt
      121 tttgctgaaa ggaggaacta tatccggatc tggcgtaata gcgaagaggc ccgcaccgat
      181 cgcccttccc aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
      241 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      301 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      361 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      421 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      481 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      541 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
      601 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
      661 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
      721 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      781 ttcaataata ttgaaaaagg aagagtatga ttgaacaaga tggattgcac gcaggttctc
      841 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
      901 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
      961 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     1021 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     1081 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     1141 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     1201 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     1261 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     1321 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     1381 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     1441 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     1501 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     1561 agcgcatcgc cttctatcgc cttcttgacg agttcttctg actgtcagac caagtttact
     1621 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1681 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1741 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1801 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1861 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1921 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1981 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     2041 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     2101 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     2161 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     2221 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     2281 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     2341 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     2401 gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta
     2461 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     2521 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2581 cgattcatta atgcaggatc tcgatcccgc gaaattaata cgactcacta tagggagacc
     2641 acaacggttt ccctctagaa tcaccgagtt tattcttgac acctgatgcg atgaatgata
     2701 taataggaaa gtactgtttt gtttaacttt aagaagggga tatacatatg cggggttctc
     2761 atcatcatca tcatcatggt atggctagca tgactggtgg acagcaaatg ggtcgggatc
     2821 tgtacgacga tgacgataag gatccggccc tgagcaacaa gttcatcggc gacgacatga
     2881 agatgaccta ccacatggac ggctgcgtga acggccacta cttcaccgtg aagggcgagg
     2941 gcaacggcaa gccctacgag ggcacccaga ccagcacctt caaggtgacc atggccaacg
     3001 gcggccccct ggccttcagc ttcgacatcc tgagcaccgt gttcaagtac ggcaaccggt
     3061 gcttcaccgc ctaccccacc agcatgcccg actacttcaa gcaggccttc cccgacggca
     3121 tgagctacga gcggaccttc acctacgagg acggcggcgt ggccaccgcc agctgggaga
     3181 tcagcctgaa gggcaactgc ttcgagcaca agagcacctt ccacggcgtg aacttccccg
     3241 ccgacggccc cgtgatggcc aagaagacca ccggctggga ccccagcttc gagaagatga
     3301 ccgtgtgcga cggcatcctg aagggcgacg tgaccgcctt cctgatgctg cagggcggcg
     3361 gcaactaccg gtgccagttc cacaccagct acaagaccaa gaagcccgtg accatgcccc
     3421 ccaaccacgt ggtggagcac cggatcgccc ggaccgacct ggacaagggc ggcaacagcg
     3481 tgcagctgac cgagcacgcc gtggcccaca tcaccagcgt ggtgcccttc taa
//
