LOCUS       proinsulin-msGFP        4371 bp DNA     circular SYN 20-JAN-2024
DEFINITION  Plasmid for bacterial expression of a proinsulin-msGFP2(C48S/C70S) 
            fusion.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4371)
  AUTHORS   Glick Lab
  TITLE     Direct Submission
  JOURNAL   Exported May 6, 2024 from SnapGene 7.1.0b
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4371
                     /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..374
                     /codon_start=1
                     /label=proinsulin
                     /note="Human proinsulin gene w/o introns"
                     /translation="MFVNQHLCGSHLVEALYLVCGERGFFYTPKTRREAEDLQVGQVEL
                     GGGPGAGSLQPLALEGSLQKRGIVEQCCTSICSLYQLENYC"
     CDS             376..1083
                     /codon_start=1
                     /label=msGFP2 (C48S C70S)
                     /translation="MDSTESLFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLK
                     FISTTGKLPVPWPTLVTTLTYGVQSFSRYPDHMKQHDFFKSAMPEGYVQERTITFKDDG
                     TYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLLE
                     FVTAAGITGGSGGS"
     terminator      1117..1211
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     terminator      1983..2069
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     misc_feature    2225..2365
                     /label=bom
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(2551..3139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number colE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3310..4101,4102..4170))
                     /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(4171..4275)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctcgagaaat cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca
       61 attgtgagcg gataacaatt tcacacagaa ttcattaaag aggagaaatt aaccatgttt
      121 gtgaaccaac acctgtgcgg ctcacacctg gtggaagctc tctacctagt gtgcggggaa
      181 cgaggcttct tctacacacc caagacccgc cgggaggcag aggacctgca ggtggggcag
      241 gtggagctgg gcgggggccc tggtgcaggc agcctgcagc ccttggccct ggaggggtcc
      301 ctgcagaagc gtggcattgt ggaacaatgc tgtaccagca tctgctccct ctaccagctg
      361 gagaactact gcaccatgga tagcactgag agcctgttca ccggggtggt gcccatcctg
      421 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgcgcggcga gggcgagggc
      481 gatgccacca acggcaagct gaccctgaag ttcatcagca ccaccggcaa gctgcccgtg
      541 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agagcttcag ccgctacccc
      601 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      661 cgcaccatca ccttcaagga cgacggcacc tacaagaccc gcgccgaggt gaagttcgag
      721 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
      781 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat caccgccgac
      841 aagcagaaga acggcatcaa ggccaacttc aagatccgcc acaacgtgga ggacggcagc
      901 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
      961 cccgacaacc actacctgag cacccagtcc aagctgagca aagaccccaa cgagaagcgc
     1021 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactggagg cagtggcggt
     1081 tcttaaagcg gccgcaagct taattagctg agcttggact cctgttgata gatccagtaa
     1141 tgacctcaga actccatctg gatttgttca gaacgctcgg ttgccgccgg gcgtttttta
     1201 ttggtgagaa tccaagctag cttggcgaga ttttcaggag ctaaggaagc taaaatggag
     1261 aaaaaaatca ctggatatac caccgttgat atatcccaat ggcatcgtaa agaacatttt
     1321 gaggcatttc agtcagttgc tcaatgtacc tataaccaga ccgttcagct ggatattacg
     1381 gcctttttaa agaccgtaaa gaaaaataag cacaagtttt atccggcctt tattcacatt
     1441 cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg caatgaaaga cggtgagctg
     1501 gtgatatggg atagtgttca cccttgttac accgttttcc atgagcaaac tgaaacgttt
     1561 tcatcgctct ggagtgaata ccacgacgat ttccggcagt ttctacacat atattcgcaa
     1621 gatgtggcgt gttacggtga aaacctggcc tatttcccta aagggtttat tgagaatatg
     1681 tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt ttgatttaaa cgtggccaat
     1741 atggacaact tcttcgcccc cgttttcacc atgcatgggc aaatattata cgcaaggcga
     1801 caaggtgctg atgccgctgg cgattcaggt tcatcatgcc gtctgtgatg gcttccatgt
     1861 cggcagaatg cttaatgaat tacaacagta ctgcgatgag tggcagggcg gggcgtaatt
     1921 tttttaaggc agttattggt gcccttaaac gcctggggta atgactctct agcttgaggc
     1981 atcaaataaa acgaaaggct cagtcgaaag actgggcctt tcgttttatc tgttgtttgt
     2041 cggtgaacgc tctcctgagt aggacaaatc cgccgctcta gagctgcctc gcgcgtttcg
     2101 gtgatgacgg tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt
     2161 aagcggatgc cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc
     2221 ggggcgcagc catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc
     2281 ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg
     2341 cgtaaggaga aaataccgca tcaggcgctc ttccgcttcc tcgctcactg actcgctgcg
     2401 ctcggtcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     2461 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     2521 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     2581 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     2641 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     2701 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     2761 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     2821 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     2881 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     2941 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     3001 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     3061 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     3121 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     3181 gaacgaaaac tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta
     3241 gatcctttta aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg
     3301 gtctgacagt taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg
     3361 ttcatccata gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc
     3421 atctggcccc agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc
     3481 agcaataaac cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc
     3541 ctccatccag tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag
     3601 tttgcgcaac gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat
     3661 ggcttcattc agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg
     3721 caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt
     3781 gttatcactc atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag
     3841 atgcttttct gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg
     3901 accgagttgc tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt
     3961 aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct
     4021 gttgagatcc agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac
     4081 tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat
     4141 aagggcgaca cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat
     4201 ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca
     4261 aataggggtt ccgcgcacat ttccccgaaa agtgccacct gctaagaaac cattattatc
     4321 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca c
//
