LOCUS       sequence-445646-        4127 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the first 4 aa of proinsulin fused to msYFP in bacteria.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4127)
  TITLE     Yellow and oxidation-resistant derivatives of a monomeric 
            superfolder GFP
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4127)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: MFVN-msYFP
FEATURES             Location/Qualifiers
     source          1..4127
                     /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)"
     primer_bind     complement(169..190)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     777..798
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      874..968
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     primer_bind     complement(1078..1097)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      1740..1826
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(1873..1895)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1981..2121
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2036..2055
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2136..2153)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2307..2895)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2387..2406)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3066..3857,3858..3926))
                     /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     3689..3708
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3927..4031)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4095..4113
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
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 tcatgccgtt 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 cctctagagc tgcctcgcgc gtttcggtga
     1861 tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt gtctgtaagc
     1921 ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg
     1981 cgcagccatg acccagtcac gtagcgatag cggagtgtat actggcttaa ctatgcggca
     2041 tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta
     2101 aggagaaaat accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     2161 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     2221 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2281 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2341 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2401 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2461 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2521 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2581 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2641 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2701 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     2761 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2821 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2881 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2941 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3001 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     3061 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     3121 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     3181 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     3241 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     3301 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     3361 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     3421 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     3481 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3541 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3601 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3661 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3721 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3781 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3841 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3901 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3961 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     4021 ggggttccgc gcacatttcc ccgaaaagtg ccacctgcta agaaaccatt attatcatga
     4081 cattaaccta taaaaatagg cgtatcacga ggccctttcg tcttcac
//
