LOCUS       sequence-444367-        5838 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  IPTG-inducible expression of C. elegans protein GLB-1 with a His 
            tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5838)
  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 5838)
  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: GLB1_SH
FEATURES             Location/Qualifiers
     source          1..5838
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1202..1393
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1387..1409)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1495..1634
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1550..1569
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1649..1666)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1820..2408)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1900..1919)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2579..3370,3371..3439))
                     /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     3202..3221
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3440..3544)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3570..4025)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3707..3728)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3919..3938
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(4062..4109)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     4105..4123
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(4176..4193)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4683..4703
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4719..4743
                     /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(4743..4762)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4744..4762)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4846..4865)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        5071..5148
                     /gene="lacI"
                     /label=lacI promoter
     CDS             join(5149..5838,1..393)
                     /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(5168..5187)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 caaatgctga atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg
       61 ctgggcgcaa tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta
      121 gtgggatacg acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa
      181 caggattttc gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc
      241 caggcggtga agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg
      301 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      361 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct
      421 cactcattag gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc
      481 cttccggtgg gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat
      541 gcaactcgta ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg
      601 ctggagcgcg acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc
      661 tcaagccttc gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc
      721 cggcatggcg gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg
      781 ctggcggggt tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag
      841 cgactgctgc tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg
      901 tgtttcgtaa agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg
      961 catcgcagga tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg
     1021 gcattgaccc tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc
     1081 ctcacaacgt tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct
     1141 ctctcgtttc atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc
     1201 agtgaccaaa caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt
     1261 aacgcttctg gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc
     1321 gcttcacgac cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg
     1381 tgaaaacctc tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc
     1441 cgggagcaga caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc
     1501 catgacccag tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag
     1561 cagattgtac tgagagtgca ccattgcggt gtgaaatacc gcacagatgc gtaaggagaa
     1621 aataccgcat caggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1681 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1741 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1801 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1861 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1921 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1981 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     2041 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     2101 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     2161 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     2221 agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt ggtatctgcg
     2281 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     2341 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     2401 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     2461 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     2521 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2581 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2641 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2701 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2761 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2821 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2881 ttgttgccat tgctgcaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2941 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     3001 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     3061 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     3121 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     3181 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     3241 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     3301 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     3361 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     3421 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     3481 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3541 cgcgcacatt tccccgaaaa gtgccaccta aattgtaagc gttaatattt tgttaaaatt
     3601 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     3661 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     3721 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     3781 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     3841 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     3901 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     3961 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     4021 cgcgtcccat tcgccaatcc ggatatagtt cctcctttca gcaaaaaacc cctcaagacc
     4081 cgtttagagg ccccaagggg ttatgctagt tattgctcag cggtggcagc agccaactca
     4141 gcttcctttc gggctttgtt agcagccgga tctcagtggt ggtggtggtg gtgctcgaga
     4201 acatgtggaa gattggaatt cttgagatgt gtctgactct cagcattgaa ttcctttcca
     4261 agtgccatcc atgcggcctt ttgttgatca ttgagacatc caacagattc aagatatcca
     4321 gtgaatacag tgaagaatgc catccacaga gctggatcca ttttgtagat acgatgacgg
     4381 ttgatggttt cacggacgta tcccttgaag acttcttcat tggtgtagac gttggcaagg
     4441 agatggcacg caagaagaat acgttggcct tgtttgtcaa atctctcact cttcttcaca
     4501 tcatctgcag tgtacttctc ggctcccttg aaatagacac gaagatccgg gaagttggtg
     4561 aagaaataac ggtagaaggc gtttccattc tcaatgtttt gagcctcagt tccaaccatt
     4621 cgtccttcaa gggacttcac acagagatca ctaatttctt gacggttcat cgacatctag
     4681 acctccttct taaagttaaa caaaattatt actagagggg aattgttatc cgctcacaat
     4741 tcccctatag tgagtcgtat taatttcgcg ggatcgagat ctcgatcctc tacgccggac
     4801 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     4861 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     4921 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat
     4981 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
     5041 agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc
     5101 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     5161 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     5221 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     5281 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     5341 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     5401 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     5461 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     5521 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     5581 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     5641 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     5701 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     5761 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     5821 ggttttcaac aaaccatg
//
