LOCUS       pHSGBlaMsfGFP        4857 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 4857)
  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..4857
                     /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..1232
                     /label=pHSGBlaM_fwd
     CDS             1446..2261
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(1451..1471)
                     /label=pHSGGFPseqrev
     primer_bind     2480..2496
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     terminator      2563..2649
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     CDS             complement(2712..3506)
                     /codon_start=1
                     /gene="bla(M)"
                     /product="beta-lactamase lacking the signal sequence"
                     /label=bla(M)
                     /note="allows cytosolic expression of beta-lactamase"
                     /translation="MHPETLVKVKDAEDQLGARVGYIELDLNSGKILESFRPEERFPMM
                     STFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYSPVTEKHLTDGMTVRELCSAAIT
                     MSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRWEPELNEAIPNDERDTTMPVAMA
                     TTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSALPAGWFIADKSGAGERGSRGII
                     AALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGASLIKHW"
     primer_bind     complement(3548..3564)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3572..3588
                     /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)."
     promoter        complement(join(3596..3602,3603..3620,3621..3626))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3641..3662
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4134..4722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4749..4767
                     /label=pHSGGFPseqfwd
     primer_bind     complement(join(4839..4857,1..20))
                     /label=pHSG298BlaM_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 tggagctttg ttgtaggtgg accagttggt gattttgaac ttttgctttg
     1261 ccacggaacg gtctgcgttg tcgggaagat gcgtgatctg atccttcaac tcagcaaaag
     1321 ttcgatttat tcaacaaagc cacgttgtgt ctcaaaatct ctgatgttac attgcacaag
     1381 ataaaaatat atcatcatga acaataaaac tgtctgctta cataaacagt aatacaaggg
     1441 gtgttatgag ccatattcaa cgggaaacgt cttgctcgaa gccgcgatta aattccaaca
     1501 tggatgctga tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga
     1561 caatctatcg attgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag
     1621 gtagcgttgc caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta
     1681 tgcctcttcc gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca
     1741 ctgcgatccc cgggaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa
     1801 atattgttga tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt
     1861 gtccttttaa cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg
     1921 gtttggttga tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct
     1981 ggaaagaaat gcataagctt ttgccattct caccggattc agtcgtcact catggtgatt
     2041 tctcacttga taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac
     2101 gagtcggaat cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt
     2161 tttctccttc attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga
     2221 ataaattgca gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt
     2281 tgtaacactg gcagagcatt acgctgactt gacgggacgg cggctttgtt gaataaatcg
     2341 cattcgccat tcaggctgcg caactgttgg gaagggcgat cggtgcgggc ctcttcgcta
     2401 ttacgccagc tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg
     2461 ttttcccagt cacgacgttg taaaacgacg gccagtgcca agcttgcatg cctgcaggtc
     2521 gactctagag gcggaatata tcccctaggt ctagggcggc ggatttgtcc tactcaggag
     2581 agcgttcacc gacaaacaac agataaaacg aaaggcccag tctttcgact gagcctttcg
     2641 ttttatttga tgcctctaga ttaattaatt aagcggccgc atcgatcggg ccctgaggcc
     2701 tgcagggatc cttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     2761 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     2821 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     2881 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     2941 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     3001 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     3061 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     3121 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     3181 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     3241 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     3301 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     3361 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     3421 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     3481 actttcacca gcgtttctgg gtgcatgggt acctttctcc gggtaccgag ctcgaattcg
     3541 taatcatgtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     3601 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     3661 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     3721 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tggcgaactt ttgctgagtt
     3781 gaaggatcag atcacgcatc ttcccgacaa cgcagaccgt tccgtggcaa agcaaaagtt
     3841 caaaatcagt aaccgtcagt gccgataagt tcaaagttaa acctggtgtt gataccaaca
     3901 ttgaaacgct gatcgaaaac gcgctgaaaa acgctgctga atgtgcgagc ttcttccgct
     3961 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4021 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4081 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4141 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4201 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4261 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4321 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4381 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4441 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4501 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4561 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4621 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4681 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4741 tctacggggt ctgacgctca gtggaacgat ccgtcgagag gtctgcctcg tgaagaaggt
     4801 gttgctgact cataccaggc ctgaatcgcc ccatcatcca gccagaaagt gagggag
//
