LOCUS       sequence-444369-        5949 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible expression of C. elegans protein GLB-21 with a His 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5949)
  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 5949)
  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: GLB21_KW
FEATURES             Location/Qualifiers
     source          1..5949
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(495..1286,1287..1355))
                     /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     1118..1137
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1356..1460)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1486..1941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1623..1644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1835..1854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(1978..2025)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     2021..2039
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(2092..2109)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             2710..2730
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2746..2770
                     /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(2770..2789)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2771..2789)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2873..2892)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        3098..3175
                     /gene="lacI"
                     /label=lacI promoter
     CDS             3176..4258
                     /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(3195..3214)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             5067..5258
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(5252..5274)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    5360..5499
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     5415..5434
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5514..5531)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(5685..5949,1..324))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5765..5784)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
       61 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
      121 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca
      181 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
      241 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
      301 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
      361 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
      421 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
      481 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
      541 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
      601 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
      661 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
      721 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
      781 aatagtttgc gcaacgttgt tgccattgct gcaggcatcg tggtgtcacg ctcgtcgttt
      841 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
      901 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
      961 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     1021 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     1081 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     1141 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     1201 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     1261 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     1321 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     1381 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     1441 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta
     1501 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1561 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1621 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1681 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1741 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1801 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1861 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1921 atgcgccgct acagggcgcg tcccattcgc caatccggat atagttcctc ctttcagcaa
     1981 aaaacccctc aagacccgtt tagaggcccc aaggggttat gctagttatt gctcagcggt
     2041 ggcagcagcc aactcagctt cctttcgggc tttgttagca gccggatctc agtggtggtg
     2101 gtggtggtgc tcgaggaatt ttgtttcttc atcatgacag ttacgacgag cctcatcgag
     2161 aaaacctttc ttcatttctc gaatcactga gctcatgaac ttttcccaac caattataat
     2221 gactgtactt gaactagaag gaaacaatga tggaacactt ttcagatcca ttgaactact
     2281 ttcagaattt ccacaatctt ttgtcactat ctccactgtc actttcttga acgtcaacca
     2341 attgtcggcg ccaaagttca ctcctctata aaaatgaatt tgtccaatac ggtatgccat
     2401 ctgatgaata tttccaattg gacagaaccc aagtgaagag actgcagtat ccaaaaagtt
     2461 ttgaattcgg tgtgcttgca attgaaattc tctgcaactg tacacatcat gattcttttc
     2521 atccacgtga attccaaaat aatccttaaa ctgtggcttc cgttcaataa gtgcgcgtag
     2581 gatgtgtttg ccaatttctc gttctttggc tctcttccac gcatctctga ctgcttgcat
     2641 ttcatcagtt gtcaattcgt acatactcct ctccgaattt cttccaaaac aatttaactt
     2701 catctagacc tccttcttaa agttaaacaa aattattact agaggggaat tgttatccgc
     2761 tcacaattcc cctatagtga gtcgtattaa tttcgcggga tcgagatctc gatcctctac
     2821 gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc
     2881 gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc
     2941 ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat
     3001 gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta
     3061 atgcaggagt cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac
     3121 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     3181 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     3241 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     3301 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     3361 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     3421 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     3481 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     3541 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     3601 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     3661 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     3721 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     3781 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     3841 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     3901 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     3961 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     4021 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     4081 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     4141 actggtgaaa agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt
     4201 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     4261 gcaacgcaat taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct
     4321 tgagagcctt caacccagtc agctccttcc ggtgggcgcg gggcatgact atcgtcgccg
     4381 cacttatgac tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg
     4441 tcattttcgg cgaggaccgc tttcgctgga gcgcgacgat gatcggcctg tcgcttgcgg
     4501 tattcggaat cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt
     4561 tcggcgagaa gcaggccatt atcgccggca tggcggcccc acgggtgcgc atgatcgtgc
     4621 tcctgtcgtt gaggacccgg ctaggctggc ggggttgcct tactggttag cagaatgaat
     4681 caccgatacg cgagcgaacg tgaagcgact gctgctgcaa aacgtctgcg acctgagcaa
     4741 caacatgaat ggtcttcggt ttccgtgttt cgtaaagtct ggaaacgcgg aagtcagcgc
     4801 cctgcaccat tatgttccgg atctgcatcg caggatgctg ctggctaccc tgtggaacac
     4861 ctacatctgt attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc
     4921 gcatccatac cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat
     4981 cagtaacccg tatcgtgagc atcctctctc gtttcatcgg tatcattacc cccatgaaca
     5041 gaaatccccc ttacacggag gcatcagtga ccaaacagga aaaaaccgcc cttaacatgg
     5101 cccgctttat cagaagccag acattaacgc ttctggagaa actcaacgag ctggacgcgg
     5161 atgaacaggc agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct
     5221 gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggagacgg
     5281 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     5341 gtgttggcgg gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata
     5401 ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa
     5461 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc
     5521 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     5581 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     5641 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     5701 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     5761 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     5821 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     5881 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     5941 cacgaaccc
//
