LOCUS       sequence-444368-        5955 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible expression of C. elegans protein GLB-10 with a His 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5955)
  AUTHORS   McAllister A, Walstrom K
  TITLE     UV-Visible Heme Absorbance Spectroscopic Characterization of 
            Caenorhabditis elegans Globin Proteins GLB-10 and GLB-31
  JOURNAL   Mendeley Data. 2025 Jan 2; V1. doi: 10.17632/td8sy4xfmp.1
REFERENCE   2  (bases 1 to 5955)
  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: "Mendeley 
            Data."; date: "2025-01-2"; volume: " V1. doi"; pages: " 
            10.17632/td8sy4xfmp."
COMMENT     Sequence Label: GLB10_MS
FEATURES             Location/Qualifiers
     source          1..5955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1286..1477
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1471..1493)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1579..1718
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1634..1653
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1733..1750)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1904..2492)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1984..2003)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2663..3454,3455..3523))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3286..3305
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3524..3628)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3654..4109)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3791..3812)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4003..4022
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4146..4193)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4189..4207
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4260..4277)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4884..4904
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4920..4944
                     /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)."
     primer_bind     complement(4944..4963)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4945..4963)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5047..5066)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5272..5349
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5350..5955,1..477)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(5369..5388)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 aaatatctca ctcgcaatca aattcagccg atagcggaac gggaaggcga ctggagtgcc
       61 atgtccggtt ttcaacaaac catgcaaatg ctgaatgagg gcatcgttcc cactgcgatg
      121 ctggttgcca acgatcagat ggcgctgggc gcaatgcgcg ccattaccga gtccgggctg
      181 cgcgttggtg cggatatctc ggtagtggga tacgacgata ccgaagacag ctcatgttat
      241 atcccgccgt taaccaccat caaacaggat tttcgcctgc tggggcaaac cagcgtggac
      301 cgcttgctgc aactctctca gggccaggcg gtgaagggca atcagctgtt gcccgtctca
      361 ctggtgaaaa gaaaaaccac cctggcgccc aatacgcaaa ccgcctctcc ccgcgcgttg
      421 gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg
      481 caacgcaatt aatgtaagtt agctcactca ttaggcaccg ggatctcgac cgatgccctt
      541 gagagccttc aacccagtca gctccttccg gtgggcgcgg ggcatgacta tcgtcgccgc
      601 acttatgact gtcttcttta tcatgcaact cgtaggacag gtgccggcag cgctctgggt
      661 cattttcggc gaggaccgct ttcgctggag cgcgacgatg atcggcctgt cgcttgcggt
      721 attcggaatc ttgcacgccc tcgctcaagc cttcgtcact ggtcccgcca ccaaacgttt
      781 cggcgagaag caggccatta tcgccggcat ggcggcccca cgggtgcgca tgatcgtgct
      841 cctgtcgttg aggacccggc taggctggcg gggttgcctt actggttagc agaatgaatc
      901 accgatacgc gagcgaacgt gaagcgactg ctgctgcaaa acgtctgcga cctgagcaac
      961 aacatgaatg gtcttcggtt tccgtgtttc gtaaagtctg gaaacgcgga agtcagcgcc
     1021 ctgcaccatt atgttccgga tctgcatcgc aggatgctgc tggctaccct gtggaacacc
     1081 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     1141 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     1201 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc ccatgaacag
     1261 aaatccccct tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc
     1321 ccgctttatc agaagccaga cattaacgct tctggagaaa ctcaacgagc tggacgcgga
     1381 tgaacaggca gacatctgtg aatcgcttca cgaccacgct gatgagcttt accgcagctg
     1441 cctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     1501 cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     1561 tgttggcggg tgtcggggcg cagccatgac ccagtcacgt agcgatagcg gagtgtatac
     1621 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccattg cggtgtgaaa
     1681 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ctcttccgct tcctcgctca
     1741 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     1801 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     1861 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     1921 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     1981 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     2041 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     2101 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     2161 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     2221 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     2281 cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac ggctacacta
     2341 gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     2401 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     2461 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     2521 ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa
     2581 ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc taaagtatat
     2641 atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct atctcagcga
     2701 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac
     2761 gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca cgctcaccgg
     2821 ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg
     2881 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2941 cgccagttaa tagtttgcgc aacgttgttg ccattgctgc aggcatcgtg gtgtcacgct
     3001 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     3061 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta
     3121 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     3181 tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     3241 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     3301 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa
     3361 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     3421 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     3481 caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc ctttttcaat
     3541 attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt
     3601 agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca cctaaattgt
     3661 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     3721 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     3781 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     3841 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     3901 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     3961 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     4021 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     4081 cgcgcttaat gcgccgctac agggcgcgtc ccattcgcca atccggatat agttcctcct
     4141 ttcagcaaaa aacccctcaa gacccgttta gaggccccaa ggggttatgc tagttattgc
     4201 tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt tgttagcagc cggatctcag
     4261 tggtggtggt ggtggtgctc gaggaatcca ttgttagagc ctcctttagt tccgccaagc
     4321 tgttgctttc gtcgatcgga atatccatta cgcatttgat gcacgattga gttaatcact
     4381 ctctgccatg ccaaaatcat ctcgttcctt ttgtcctcgt ctggtatata agctgaaagt
     4441 gtctcatcaa tcttctctgc catgcaaacc aagaatatat cccagaaatg tgaattgaat
     4501 ccacgagaaa tgaaatcaac atgccgggct cccaattgtt cacaagcttc tctcacttga
     4561 tcatcattaa gctcatatgt gatgatcaat ttgtagaaga atgcttggat tgttgaagaa
     4621 agacccaaaa atgctgcatt ttgcattggg ctctcatttt ctgcgattcc aaatgccagt
     4681 ttgattgaag tggatcgtgt agccgatttg tgccagattt ctgtgaagag ctcagggcat
     4741 tgagcaagaa ctcttggcca ttcttgaatc accaatttct ttgttttagc attcaatggt
     4801 ccaacttgtg ctgagccaat tccatttcca accactggtt ccttctttcc gcggatcgtt
     4861 gacaacaagc ccatcatcta gacctccttc ttaaagttaa acaaaattat tactagaggg
     4921 gaattgttat ccgctcacaa ttcccctata gtgagtcgta ttaatttcgc gggatcgaga
     4981 tctcgatcct ctacgccgga cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg
     5041 ctggcgccta tatcgccgac atcaccgatg gggaagatcg ggctcgccac ttcgggctca
     5101 tgagcgcttg tttcggcgtg ggtatggtgg caggccccgt ggccggggga ctgttgggcg
     5161 ccatctcctt gcatgcacca ttccttgcgg cggcggtgct caacggcctc aacctactac
     5221 tgggctgctt cctaatgcag gagtcgcata agggagagcg tcgagatccc ggacaccatc
     5281 gaatggcgca aaacctttcg cggtatggca tgatagcgcc cggaagagag tcaattcagg
     5341 gtggtgaatg tgaaaccagt aacgttatac gatgtcgcag agtatgccgg tgtctcttat
     5401 cagaccgttt cccgcgtggt gaaccaggcc agccacgttt ctgcgaaaac gcgggaaaaa
     5461 gtggaagcgg cgatggcgga gctgaattac attcccaacc gcgtggcaca acaactggcg
     5521 ggcaaacagt cgttgctgat tggcgttgcc acctccagtc tggccctgca cgcgccgtcg
     5581 caaattgtcg cggcgattaa atctcgcgcc gatcaactgg gtgccagcgt ggtggtgtcg
     5641 atggtagaac gaagcggcgt cgaagcctgt aaagcggcgg tgcacaatct tctcgcgcaa
     5701 cgcgtcagtg ggctgatcat taactatccg ctggatgacc aggatgccat tgctgtggaa
     5761 gctgcctgca ctaatgttcc ggcgttattt cttgatgtct ctgaccagac acccatcaac
     5821 agtattattt tctcccatga agacggtacg cgactgggcg tggagcatct ggtcgcattg
     5881 ggtcaccagc aaatcgcgct gttagcgggc ccattaagtt ctgtctcggc gcgtctgcgt
     5941 ctggctggct ggcat
//
