LOCUS       sequence-444370-        5868 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG inducible expression of C. elegans protein GLB-31 with a His 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5868)
  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 5868)
  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: GLB31_CN
FEATURES             Location/Qualifiers
     source          1..5868
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      413..1001
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     902..921
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1155..1172
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1187..1326
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1252..1271)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1412..1434
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1428..1619)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2428..3510)
                     /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     3472..3491
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(3511..3588)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     3794..3813
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     3897..3916
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3897..3915
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    3916..3940
                     /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             3956..3976
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4496..4513
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4566..4584)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4580..4627
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4664..5119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4751..4770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4961..4982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        5145..5249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5250..5318,5319..5868,1..242)
                     /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     complement(5468..5487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
       61 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
      121 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
      181 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
      241 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
      301 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
      361 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
      421 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
      481 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
      541 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
      601 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
      661 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
      721 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
      781 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
      841 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
      901 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
      961 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     1021 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     1081 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     1141 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg atgcggtatt
     1201 ttctccttac gcatctgtgc ggtatttcac accgcaatgg tgcactctca gtacaatctg
     1261 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     1321 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     1381 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     1441 ccgtcatcac cgaaacgcgc gaggcagctg cggtaaagct catcagcgtg gtcgtgaagc
     1501 gattcacaga tgtctgcctg ttcatccgcg tccagctcgt tgagtttctc cagaagcgtt
     1561 aatgtctggc ttctgataaa gcgggccatg ttaagggcgg ttttttcctg tttggtcact
     1621 gatgcctccg tgtaaggggg atttctgttc atgggggtaa tgataccgat gaaacgagag
     1681 aggatgctca cgatacgggt tactgatgat gaacatgccc ggttactgga acgttgtgag
     1741 ggtaaacaac tggcggtatg gatgcggcgg gaccagagaa aaatcactca gggtcaatgc
     1801 cagcgcttcg ttaatacaga tgtaggtgtt ccacagggta gccagcagca tcctgcgatg
     1861 cagatccgga acataatggt gcagggcgct gacttccgcg tttccagact ttacgaaaca
     1921 cggaaaccga agaccattca tgttgttgct caggtcgcag acgttttgca gcagcagtcg
     1981 cttcacgttc gctcgcgtat cggtgattca ttctgctaac cagtaaggca accccgccag
     2041 cctagccggg tcctcaacga caggagcacg atcatgcgca cccgtggggc cgccatgccg
     2101 gcgataatgg cctgcttctc gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga
     2161 gcgagggcgt gcaagattcc gaataccgca agcgacaggc cgatcatcgt cgcgctccag
     2221 cgaaagcggt cctcgccgaa aatgacccag agcgctgccg gcacctgtcc tacgagttgc
     2281 atgataaaga agacagtcat aagtgcggcg acgatagtca tgccccgcgc ccaccggaag
     2341 gagctgactg ggttgaaggc tctcaagggc atcggtcgag atcccggtgc ctaatgagtg
     2401 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2461 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     2521 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     2581 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     2641 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     2701 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     2761 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     2821 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     2881 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     2941 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     3001 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     3061 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     3121 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     3181 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     3241 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     3301 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     3361 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     3421 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     3481 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     3541 gcgctatcat gccataccgc gaaaggtttt gcgccattcg atggtgtccg ggatctcgac
     3601 gctctccctt atgcgactcc tgcattagga agcagcccag tagtaggttg aggccgttga
     3661 gcaccgccgc cgcaaggaat ggtgcatgca aggagatggc gcccaacagt cccccggcca
     3721 cggggcctgc caccataccc acgccgaaac aagcgctcat gagcccgaag tggcgagccc
     3781 gatcttcccc atcggtgatg tcggcgatat aggcgccagc aaccgcacct gtggcgccgg
     3841 tgatgccggc cacgatgcgt ccggcgtaga ggatcgagat ctcgatcccg cgaaattaat
     3901 acgactcact ataggggaat tgtgagcgga taacaattcc cctctagtaa taattttgtt
     3961 taactttaag aaggaggtct agatgggcat aattttggcg tatttccgac gtcccaagct
     4021 tgatatcgac agggttcggt cggtctggat ggatcatatc aatggaaacg atcagtactt
     4081 ccaagaggta atccatagaa tatgcaagcg aaacgaggga atccggtgtg caatgctggc
     4141 tccgaacgcg cagcacgccg agtcggtggc cgaggaggat tttgtgctct caaatattgc
     4201 agataggatc tcacagtttt ttcatcaatt ggttgaggat gacgtgctta tggacacggt
     4261 agaactgaaa aaagcctgct atgaccttgg aagacaacat tcttcgtatt cgaaacaaca
     4321 atttaagatg tcctactggg aagagtttac attgacaatg atgggtgttc tggagcaaaa
     4381 ctacccggaa accacgaaag aagagcaaaa agcgtggctt cactttttga gatttgtgaa
     4441 tgagaatatg ttggatggat atttagatgc gatttctcgt agtaataaac tcgagcacca
     4501 ccaccaccac cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc
     4561 tgccaccgct gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg
     4621 ttttttgctg aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc
     4681 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     4741 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     4801 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     4861 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     4921 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     4981 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     5041 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5101 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     5161 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     5221 tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt gtcgccctta
     5281 ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg ctggtgaaag
     5341 taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg gatctcaaca
     5401 gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg agcactttta
     5461 aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag caactcggtc
     5521 gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca gaaaagcatc
     5581 ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg agtgataaca
     5641 ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc gcttttttgc
     5701 acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg aatgaagcca
     5761 taccaaacga cgagcgtgac accacgatgc ctgcagcaat ggcaacaacg ttgcgcaaac
     5821 tattaactgg cgaactactt actctagctt cccggcaaca attaatag
//
