pUC-GFP-AT

LOCUS       pUC-GFP-AT               6475 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  ""Burdensome"" GFP expressing plasmid carrying axe/txe 
            toxin-antitoxin for plasmid stabilisation.
ACCESSION   .
VERSION     .
KEYWORDS    pUC-GFP-AT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6475)
  AUTHORS   Fedorec AJH, Ozdemir T, Doshi A, Ho YK, Rosa L, Rutter J, Velazquez 
            O, Pinheiro VB, Danino T, Barnes CP
  TITLE     Two New Plasmid Post-segregational Killing Mechanisms for the 
            Implementation of Synthetic Gene Networks in Escherichia coli.
  JOURNAL   iScience. 2019 Apr 26;14:323-334. doi: 10.1016/j.isci.2019.03.019. 
            Epub 2019 Mar 22.
  PUBMED    30954530
REFERENCE   2  (bases 1 to 6475)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6475
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    58..129
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     terminator      134..180
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    198..246
                     /note="synthetic polyadenylation signal"
     polyA_signal    2253..2374
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2264..2283)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2318..2337
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     terminator      2489..2535
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2565..2701
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2746..2817
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     rep_origin      complement(3043..3631)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3123..3142)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        4251..4319
                     /label=recA (Delta-LexA) promoter
                     /note="constitutively active E. coli recA promoter lacking 
                     the LexA binding site (Brent and Ptashne, 1981)"
     CDS             4336..5046
                     /codon_start=1
                     /product="green fluorescent protein with peak excitation at
                     510 nm and peak emission at 521 nm"
                     /label=daGFP
                     /translation="MTALTEGAKLFEKEIPYITELEGDVEGMKFIIKGEGTGDATTGTI
                     KAKYICTTGDLPVPWATLVSTLSYGVQCFAKYPSHIKDFFKSAMPEGYTQERTISFEGD
                     GVYKTRAMVTYERGSIYNRVTLTGENFKKDGHILRKNVAFQCPPSILYILPDTVNNGIR
                     VEFNQAYDIEGVTEKLVTKCSQMNRPLAGSAAVHIPRYHHITYHTKLSKDRDERRDHMC
                     LVEVVKAVDLDTYQ"
     terminator      5053..5188
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     CDS             5445..6239
                     /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(5499..5518)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6109..6128
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     terminator      6252..6388
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
ORIGIN
        1 atcgaatctc gtcagctatc gtcagcttac ctttttggca gcgatcgcgg ctcccgacat
       61 cttggaccat tagctccaca ggtatcttct tccctctagt ggtcataaca gcagcttcag
      121 ctacctctca attcaaaaaa cccctcaaga cccgtttaga ggccccaagg ggttatgcta
      181 tcaatcgttg cgttacacac acaaaaaacc aacacacatc catcttcgat ggatagcgat
      241 tttattatct aactgctgat cgagtgtagc cagatctttg tcgatcctac catccactcg
      301 acacacccgc cagcggccgc tgccaagctt ccgagctcga tacttctttc aaattcaagt
      361 taaatttata ttccattact tctcctccaa tatataaaaa ttacttaact gtatccaaat
      421 ttaatatgta aataaatcat taatttttag ctaaattcta tgagaagttt atttttgttg
      481 tttttgctaa ttgactaaaa ctagtacaaa aaatttaaaa tagattatta aacctattgc
      541 ttattctatt cagtcatata tataagcgtc atcctttagt aactaacatc ccacactaca
      601 atagtattat aacatatcta ttcaacttca aattgttgaa atttaagcca tttatattta
      661 cttttcctca actttagaaa tgattttact aaccttatat ccgtaatacg cgtaacaaac
      721 acattcatta ataacaatca acactaagct atattagctt tttaaatgac tagggtaaat
      781 aaaataagga tcaaaaactt ttaagtttct gaccctttcc ttacttccga ttggttaata
      841 gtgatctttt gcagaataaa taaatatcgt ttcattttca actctatata tcagtctatg
      901 ttcatctgta attcttctgg accattttcc agataaatca tgctttaatg gctcaggttt
      961 tcctaatcca gcaaaggggg aacgatcgat atcttttatt aacttgttaa tcttttttat
     1021 attgcttttg tttccttgct catgccaata aagataatca tcccaagcat catcagacca
     1081 agccttaatc atcagattca acctcgatta agtcatgtgt tttaaatgca cctttggaga
     1141 attgttcatc tcctcgacga attttttcca tgacgtaatt attagaaagt gttctcaacg
     1201 tttcttgcat agaatcataa tctctttttg ataatacaac aactgtatct tctacatctt
     1261 tacttgttac aataagtgtt tcagcatcct cattaacttg tttcatataa ctacgtaaat
     1321 tttggcggaa atttgaataa gctactgctt ccattccttt caccccttat ttatttctat
     1381 aaaacaattg tacattatat tgtacaatta agcaaatgat taatattcta acttcttata
     1441 attaaaagtg ctatcacttt tagtttttag gataaaaggt gacgtcactt tttgagtgct
     1501 ggtacctcaa atcctgcttt ttcttgggca atcaataaat ctaacgcaat acacataatc
     1561 ctggattcac ttttgccgta cttctgcttt agttgctgca attctttttg tctcttttca
     1621 tccaacgtaa cgggtaaccg tttatttcct attgcaattg gcattagtcg cttatggatc
     1681 ctgaacttga ccgtaaacga atgccatatc aacgatttga gaggaaaaaa ggcaataatc
     1741 atttataccg aaaagtgaga atgtgtgaaa taaggacaaa aaacctagta actaagggaa
     1801 tggttcttat ttcgcaccaa tcgctccaac cgcccccgcc gcttaggtgg tgcgaaataa
     1861 gcgggcaaca cccgataacc taagagaaag aaggagatcc atatgaataa aatcattcct
     1921 ctattgtcag tcctatttgt tggcgaaata cttatgcgcc atgctgcagc tgtgcattta
     1981 aacttcgtag tatttttggc aatactggta ttttgtttgg tcttttggcg ctttgtatta
     2041 gcccttgtga gtctactatt caagatgatt gcagccatat ttttcatagg gttatttcga
     2101 tgaaaggaga gagtatcgga tcccccgggc tcgaggtttc tagaagttgt ctcctcctgc
     2161 actgactgac tgatacaatc gatttctgga tccgcaggcc tctgctagct tgactgactg
     2221 agatacagcg taccttcagc tcacagacat gataagatac attgatgagt ttggacaaac
     2281 cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt
     2341 atttgtaacc attataagct gcaataaaca agttaacaac aacaattgca ttcattttat
     2401 gtttcaggtt cagggggagg tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg
     2461 tggtattggc ccatctctat cggtatcgta gcataacccc ttggggcctc taaacgggtc
     2521 ttgaggggtt ttttgtgccc ctcgggccgg attgctatct accggcattg gcgcagaaaa
     2581 aaatgcctga tgcgacgctg cgcgtcttat actcccacat atgccagatt cagcaacgga
     2641 tacggcttcc ccaacttgcc cacttccata cgtgtcctcc ttaccagaaa tttatcctta
     2701 aggtcgtcag ctatcctgca ggcgatctct cgatttcgat caagacattc ctttaatggt
     2761 cttttctgga caccactagg ggtcagaagt agttcatcaa actttcttcc ctccctaatc
     2821 tcattggtta ccttgggcta tcgaaactta attaaccagt caagtcagct acttggcgag
     2881 atcgacttgt ctgggtttcg actacgctca gaattgcgtc agtcaagttc gatctggtcc
     2941 ttgctattgc acccgttctc cgattacgag tttcatttaa atcatgtgag caaaaggcca
     3001 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     3061 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     3121 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     3181 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     3241 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     3301 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     3361 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     3421 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     3481 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     3541 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     3601 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     3661 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     3721 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     3781 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     3841 ctgtctattt cgttcatcca tagttgcatt taaatttccg aactctccaa ggccctcgtc
     3901 ggaaaatctt caaacctttc gtccgatcca tcttgcaggc tacctctcga acgaactatc
     3961 gcaagtctct tggccggcct tgcgccttgg ctattgcttg gcagcgccta tcgccaggta
     4021 ttactccaat cccgaatatc cgagatcggg atcacccgag agaagttcaa cctacatcct
     4081 caatcccgat ctatccgaga tccgaggaat atcgaaatcg gggcgcgcca agctgttgtg
     4141 accgcttgct ctagccagct atcgagttgt gaaccgatcc atctagcaat tggtctcgat
     4201 ctagcgatag gcttcgatct agctatgtag aaacgccgtg tgctcgatcg cttgataagg
     4261 tccacgtagc tgctataatt gcttcaacag aacatattga ctatccggta ttacccggca
     4321 aaggaggtag ccaacatgac agctctgaca gagggagcga aactgttcga gaaggagatc
     4381 ccgtacatca cagagttgga aggagatgtc gaagggatga aattcatcat caagggagaa
     4441 ggaacgggtg acgcaacgac cggaacgatc aaggccaaat acatttgtac cacgggcgat
     4501 ttgcctgtcc cctgggcgac gcttgtaagc acgctctcgt atggtgtcca gtgcttcgcg
     4561 aaatatccat cgcacattaa ggactttttc aagtcggcca tgccagaagg ttacacacaa
     4621 gaacgaacca tctcctttga gggggacgga gtgtataaga cacgcgcgat ggtaacgtac
     4681 gagcgcgggt ccatctacaa cagggtaact cttactgggg agaactttaa gaaggacggg
     4741 catatcttgc ggaaaaacgt ggcatttcaa tgtccgccct cgattctgta tattctcccg
     4801 gacacggtga ataatgggat cagagtggag ttcaaccagg catacgatat tgagggtgtg
     4861 actgaaaagc tcgtcaccaa atgcagccag atgaatcgcc cccttgcggg atcagcagcc
     4921 gtccatatcc cccggtatca ccatattacc taccacacaa aactctcaaa agacagagat
     4981 gagagaaggg accatatgtg cctggtcgaa gtagtgaagg cggtggatct tgatacatac
     5041 cagtaggcta gccattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     5101 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     5161 gtcctcctta ccagaaattt atccttaagg gcgcgcctgg tgtaccgaga acgatcctct
     5221 cagtgcgagt ctcgacgatc catatcgttg cttggcagtc agccagtcgg aatccagctt
     5281 gggacccagg aagtccaatc gtcagatatt gtactcaagc ctggtcacgg cagcgtaccg
     5341 atctgtttaa acctagatat tgatagtctg atcggtcaac gtataatcga gtcctagctt
     5401 ttgcaaacat ctatcaagag acaggatcag caggaggctt tcgcatgatt gaacaagatg
     5461 gattgcacgc aggttctccg gcggcttggg tggagaggct attcggctat gactgggcac
     5521 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgtccgg
     5581 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     5641 ggctatcgtg gctggcgacg acgggcgttc cttgcgcggc tgtgctcgac gttgtcactg
     5701 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5761 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5821 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5881 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5941 cgccagccga actgttcgcc aggctcaagg cgtctatgcc cgacggcgag gatctcgtcg
     6001 tgacccacgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     6061 tcatcgactg tggccgtctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6121 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttccttgtg ctttacggta
     6181 tcgccgcgcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgac
     6241 cgattctagg tgcattggcg cagaaaaaaa tgcctgatgc gacgctgcgc gtcttatact
     6301 cccacatatg ccagattcag caacggatac ggcttcccca acttgcccac ttccatacgt
     6361 gtcctcctta ccagaaattt atccttaagg tcgtttaaac tcgactctgg ctctatcgaa
     6421 tctccgtcgt ttcgagctta cgcgaacagc cgtggcgctc atttgctcgt cgggc
//