LOCUS       pBlasfGFP        4568 bp DNA     circular SYN 17-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4568)
  AUTHORS   Helena Ma
  TITLE     Direct Submission
  JOURNAL   Exported Jul 9, 2024 from SnapGene Viewer 7.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4568
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     RBS             289..300
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             307..1020
                     /codon_start=1
                     /product="GFP variant that folds robustly even when fused 
                     to poorly folded proteins (Pedelacq et al., 2006)"
                     /label=superfolder GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     terminator      1032..1103
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1119..1146
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     1193..1231
                     /label=pBla_fwd
     primer_bind     complement(1340..1359)
                     /label=pBlaGFPseqrev
     CDS             1357..2151
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     terminator      2182..2276
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2390..2935
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     terminator      3105..3191
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(join(3254..4045,4046..4114))
                     /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"
     RBS             4122..4133
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     protein_bind    4152..4168
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    4182..4201
                     /label=lac operator (symmetric)
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG). The 
                     symmetric lac operator was optimized for tight binding of 
                     lac repressor."
     CDS             complement(4378..4407)
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     primer_bind     4413..4433
                     /label=pBlaGFPseqfwd
     primer_bind     complement(join(4549..4568,1..20))
                     /label=pBla_rev
ORIGIN
        1 cgagctcggt accaaattcc agaaaagagg cctcccgaaa ggggggcctt ttttcgtttt
       61 ggtccgctgc caaaccagat gtcaacacag ctacaacgtt tatagctagc tcagcccttg
      121 gtacaatgct agctgagacg gaaagtgaaa cgtgatttca tgcgtcattt tgaacatttt
      181 gtaaatctta tttaataatg tgtgcggcaa ttcacattta atttatgaat gttttcttaa
      241 catcgcggca actcaagaaa cggcaggttc ggatcttagc tactagagaa agaggagaaa
      301 tactagatgc gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg
      361 gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact
      421 aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt tccttggccg
      481 actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccatatg
      541 aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt
      601 tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc
      661 ctggtaaacc gcattgagct gaaaggcatt gactttaaag aggacggcaa tatcctgggc
      721 cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa
      781 aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg
      841 gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat
      901 cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcatatg
      961 gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa
     1021 tgaccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     1081 gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg ggtgggcctt
     1141 tctgcgttta tacgtctcag ttttagagct agaaatagca agttaaaata aggctagtcc
     1201 gttatcaact tgcagacgtc ggaattgcca gctggggcgc cctctggtaa ggttgggaag
     1261 ccctgcaaag taaactggat ggctttcttg ccgccaagga tctgatggcg caggggatca
     1321 agatctgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     1381 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     1441 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     1501 gtcaagaccg acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg
     1561 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     1621 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     1681 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     1741 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     1801 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     1861 gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat
     1921 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     1981 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     2041 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     2101 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     2161 tggggttcga gagctcgctt ggactcctgt tgatagatcc agtaatgacc tcagaactcc
     2221 atctggattt gttcagaacg ctcggttgcc gccgggcgtt ttttattggt gagaatccaa
     2281 gcactagtaa caacttatat cgtatggggc tgacttcagg tgctacattt gaagagataa
     2341 attgcactga aatctagtaa tattttatct gattaataag atgatcttct tgagatcgtt
     2401 ttggtctgcg cgtaatctct tgctctgaaa acgaaaaaac cgccttgcag ggcggttttt
     2461 cgaaggttct ctgagctacc aactctttga accgaggtaa ctggcttgga ggagcgcagt
     2521 caccaaaact tgtcctttca gtttagcctt aaccggcgca tgacttcaag actaactcct
     2581 ctaaatcaat taccagtggc tgctgccagt ggtgcttttg catgtctttc cgggttggac
     2641 tcaagacgat agttaccgga taaggcgcag cggtcggact gaacgggggg ttcgtgcata
     2701 cagtccagct tggagcgaac tgcctacccg gaactgagtg tcaggcgtgg aatgagacaa
     2761 acgcggccat aacagcggaa tgacaccggt aaaccgaaag gcaggaacag gagagcgcac
     2821 gagggagccg ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacca
     2881 ctgatttgag cgtcagattt cgtgatgctt gtcagggggg cggagcctat ggaaaaacgg
     2941 ctttgccgcg gccctctcac ttccctgtta agtatcttcc tggcatcttc caggaaatct
     3001 ccgccccgtt cgtaagccat ttccgctcgc cgcagtcgaa cgaccgagcg tagcgagtca
     3061 gtgagcgagg aagcggaata tatcccctag gtctagggcg gcggatttgt cctactcagg
     3121 agagcgttca ccgacaaaca acagataaaa cgaaaggccc agtctttcga ctgagccttt
     3181 cgttttattt gatgcctcta gattaattaa ttaagcggcc gcatcgatcg ggccctgagg
     3241 cctgcaggga tccttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     3301 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     3361 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     3421 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     3481 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     3541 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     3601 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     3661 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     3721 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     3781 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     3841 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     3901 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     3961 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     4021 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     4081 gaataagggc gacacggaaa tgttgaatac tcatgggtac ctttctcctc tttaatgaat
     4141 tctgtgtgaa attgttatcc gctcacaatt gaatctatca taattgtgag cgctcacaat
     4201 tgtaaaggtt agatccgcta atcttatgga taaaaatgct atgctcgagt cgacagttca
     4261 taggtgattg ctcaggacat ttctgttaga aggaatcgtt ttccttactt ttccttacgc
     4321 acaagagttc cgtagctgtt caagtttgtg tttcaactgt tctcgtcgtt tccgcaacaa
     4381 gtcctcttca gaaatgagct tttgctcctc tgcttggacg gacaggatgt atgctgtggc
     4441 ttttttaagg ataactacct tgggggcctt ttcattgttt tccaactccg ggatctggtc
     4501 acgcagggca aaaaagctcc gttttagctc gttcctcctc tggcgctcca agacgttgtg
     4561 tgttcgcc
//
