LOCUS       MFVN-msYFP        4128 bp DNA     circular SYN 16-JAN-2024
DEFINITION  Plasmid for bacterial expression of the first four amino acids of 
            proinsulin fused to msYFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4128)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported Apr 30, 2024 from SnapGene 7.1.0b
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4128
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        join(10..24,25..30,31..47,48..54)
                     /label=T5 promoter
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
     protein_bind    30..46
                     /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    62..78
                     /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)."
     RBS             97..108
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             115..126
                     /codon_start=1
                     /label=MFVN
                     /note="Human proinsulin gene w/o introns"
                     /translation="MFVN"
     CDS             127..843
                     /codon_start=1
                     /label=msYFP
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     LICTTGKLPVPWPTLVTTLGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDHMVLLE
                     FVTAAGITHGSSGSSG"
     terminator      874..968
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1740..1826
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    1982..2122
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2308..2896)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3067..3858,3859..3927))
                     /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"
     promoter        complement(3928..4032)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatgttt
      121 gtgaacatgg atagcactga gagcctgttc accggggtgg tgcccatcct ggtcgagctg
      181 gacggcgacg taaacggcca caagttcagc gtgcgcggcg agggcgaggg cgatgccacc
      241 aacggcaagc tgaccctgaa gctgatctgc accaccggca agctgcccgt gccctggccc
      301 accctcgtga ccaccctggg ctacggcgtg cagtgcttcg cccgctaccc cgaccacatg
      361 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      421 accttcaagg acgacggcac ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
      481 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
      541 cacaagctgg agtacaactt caacagccac aacgtctata tcaccgccga caagcagaag
      601 aacggcatca aggccaactt caagatccgc cacaacgtgg aggacggcgg cgtgcagctc
      661 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
      721 cactacctga gctaccagtc caagctgagc aaagacccca acgagaagcg cgatcacatg
      781 gtcctgctgg agttcgtgac cgccgccggg atcactcacg gcagctccgg ctccagcggg
      841 taaagcggcc gcaagcttaa ttagctgagc ttggactcct gttgatagat ccagtaatga
      901 cctcagaact ccatctggat ttgttcagaa cgctcggttg ccgccgggcg ttttttattg
      961 gtgagaatcc aagctagctt ggcgagattt tcaggagcta aggaagctaa aatggagaaa
     1021 aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga acattttgag
     1081 gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga tattacggcc
     1141 tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat tcacattctt
     1201 gcccgcctga tgaatgctca tccggaattt cgtatggcaa tgaaagacgg tgagctggtg
     1261 atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga aacgttttca
     1321 tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata ttcgcaagat
     1381 gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga gaatatgttt
     1441 ttcgtctcag ccaatccctg ggtgagtttc accagttttg atttaaacgt ggccaatatg
     1501 gacaacttct tcgcccccgt tttcaccatg catgggcaaa tattatacgc aaggcgacaa
     1561 ggtgctgatg ccgctggcga ttcaggttca tcatgccgtc tgtgatggct tccatgtcgg
     1621 cagaatgctt aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaattttt
     1681 ttaaggcagt tattggtgcc cttaaacgcc tggggtaatg actctctagc ttgaggcatc
     1741 aaataaaacg aaaggctcag tcgaaagact gggcctttcg ttttatctgt tgtttgtcgg
     1801 tgaacgctct cctgagtagg acaaatccgc cgctctagag ctgcctcgcg cgtttcggtg
     1861 atgacggtga aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag
     1921 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     1981 gcgcagccat gacccagtca cgtagcgata gcggagtgta tactggctta actatgcggc
     2041 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     2101 aaggagaaaa taccgcatca ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     2161 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     2221 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     2281 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     2341 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     2401 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     2461 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta
     2521 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     2581 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     2641 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     2701 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     2761 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     2821 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     2881 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     2941 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     3001 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     3061 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     3121 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     3181 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     3241 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     3301 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     3361 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     3421 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     3481 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     3541 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     3601 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     3661 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     3721 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     3781 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     3841 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     3901 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     3961 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     4021 aggggttccg cgcacatttc cccgaaaagt gccacctgct aagaaaccat tattatcatg
     4081 acattaacct ataaaaatag gcgtatcacg aggccctttc gtcttcac
//
