LOCUS       pCN-mBFP                3432 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Encodes a NADPH dependent blue fluorescent protein expressed under a
            strong promoter and strong RBS without His-tagged. .
ACCESSION   .
VERSION     .
KEYWORDS    pCN-mBFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3432)
  AUTHORS   Ng CY, Farasat I, Maranas CD, Salis HM
  TITLE     Rational design of a synthetic Entner-Doudoroff pathway for improved
            and controllable NADPH regeneration.
  JOURNAL   Metab Eng. 2015 May;29:86-96. doi: 10.1016/j.ymben.2015.03.001. Epub
            2015 Mar 10.
  PUBMED    25769287
REFERENCE   2  (bases 1 to 3432)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            //gt12365.cn
FEATURES             Location/Qualifiers
     source          1..3432
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      989..1083
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1159..1181)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1267..1407
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1322..1341
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1422..1439)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1593..2181)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1673..1692)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(2363..3157)
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFNDCGRLGVADRYQDIA
                     LDTRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     3084..3103
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3392..3410
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
ORIGIN
        1 ttcttgagca cagctaacac cacgtcgtcc ctatctgctg ccctaggtct atgagtggtt
       61 gctggataac gacgtctttt ttttgacagc tagctcagtc ctaggtataa tatattcagg
      121 gagaccacag gatccacggt ttccctctac aaataatttt gtttaacttt tactagagga
      181 attcttaaag acccaggacg atttaaggag gagaaaagac atgcagaatc tgaacggcaa
      241 agtggctttc gtgaccggcg gcagccgcgg catcggcgcg gcgatcgtcc gccgcttggc
      301 ggcggacggc gccgacatcg cgttcaccta tgtcagcgcc tcgtcgaaaa acgtggccac
      361 cgccctggtg caagaactcg aggccaaggg ccgccgcgct cgcgccatcc aggcggactc
      421 ggcggatccg gcccaggtgc ggcaggcggt cgagcaggcc atcgtgcaac tggggccggt
      481 ggacgtgctg gtgaacaacg ccggcatctt cctggccggc cccttgggcg aggtgacgct
      541 ggacgactac gaacgcacga tgaacatcaa tgtgcgcgcg cctttcgtgg ccatccaggc
      601 cgcgcaggcc tcgatgccgg acggcggccg catcatcaac atcggcagct gcctggcgga
      661 acgcgccggc cgagccgggg taacgctgta tgccgccagc aagtcggcgc tgctgggcat
      721 gacgcgcggc ctggcgcgcg acctgggcgc gcgcggcatc accgccaacg tcgtgcaccc
      781 gggcccgatc gacaccgaca tgaatcccgc agatggcgaa cgctcgggcg aactggtggc
      841 cgtgctgtcc ttgcctcatt acggcgaggt gcgcgacatc gccggcatgg tggctttcct
      901 ggccgggccg gatgggcgct acgtgaccgg tgcgagtctg gcggtggacg gcggcttcgc
      961 cgcttgaaag cttaattagc tgagcttgga ctcctgttga tagatccagt aatgacctca
     1021 gaactccatc tggatttgtt cagaacgctc ggttgccgcc gggcgttttt tattggtgag
     1081 aatccaagct agcttggcga gattttcagg agctaaggtc tagagctgcc tcgcgcgttt
     1141 cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct
     1201 gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg
     1261 tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg gcttaactat
     1321 gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga
     1381 tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac tgactcgctg
     1441 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     1501 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     1561 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     1621 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     1681 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     1741 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     1801 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     1861 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     1921 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     1981 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta
     2041 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     2101 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     2161 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     2221 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     2281 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     2341 tggtctgaca gttaccgagc tctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     2401 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     2461 caagctcttc agcaatatca cgggtatcca acgctatgtc ctgatagcgg tccgccacac
     2521 ccagccggcc acagtcgttg aatccagaaa agcggccatt ttccaccatg atattcggca
     2581 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgcgc gccttgagcc
     2641 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     2701 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     2761 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     2821 ctttctcggc aggagcaagg tgggatgaca ggagatcctg ccccggcact tcgcccaata
     2881 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     2941 tcgtggccag ccacgatagc cgcgctgcct cgtcctgcag ttcattcagg gcaccggaca
     3001 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     3061 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     3121 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     3181 cttgatcaga tcttgatccc ctgcgccatc agatccttgg cggcaagaaa gccatccagt
     3241 ttactttgca gggcttccca accttaccag agggcgcccc agctggcaat tccggttcgc
     3301 ttgctgtcca taaaaccgcc cagtctagct atcgggtacc gtgccacctg acgtctaaga
     3361 aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct
     3421 tcacctgtcg ac
//