LOCUS       pHSG298BlasfGFP        4924 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 4924)
  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..4924
                     /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"
     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     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(join(2712..3503,3504..3572))
                     /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"
     primer_bind     complement(3614..3630)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3638..3654
                     /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(3662..3668,3669..3686,3687..3692))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3707..3728
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4200..4788)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     3541 ataagggcga cacggaaatg ttgaatactc atgggtacct ttctccgggt accgagctcg
     3601 aattcgtaat catgtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3661 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3721 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3781 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggc gaacttttgc
     3841 tgagttgaag gatcagatca cgcatcttcc cgacaacgca gaccgttccg tggcaaagca
     3901 aaagttcaaa atcagtaacc gtcagtgccg ataagttcaa agttaaacct ggtgttgata
     3961 ccaacattga aacgctgatc gaaaacgcgc tgaaaaacgc tgctgaatgt gcgagcttct
     4021 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4081 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4141 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4201 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4261 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4321 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4381 gtggcgcttt ctcaatgctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4441 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4501 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4561 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     4621 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     4681 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4741 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4801 atcttttcta cggggtctga cgctcagtgg aacgatccgt cgagaggtct gcctcgtgaa
     4861 gaaggtgttg ctgactcata ccaggcctga atcgccccat catccagcca gaaagtgagg
     4921 gagc
//
