LOCUS       sequence-148282-        7803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses the N-terminal fragment of manduca sexta soluble guanylyl 
            cyclase subunits alpha and beta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7803)
  AUTHORS   Fritz BG, Roberts SA, Ahmed A, Breci L, Li W, Weichsel A, Brailey 
            JL, Wysocki VH, Tama F, Montfort WR
  TITLE     Molecular model of a soluble guanylyl cyclase fragment determined by
            small-angle X-ray scattering and chemical cross-linking.
  JOURNAL   Biochemistry. 2013 Mar 5;52(9):1568-82. doi: 10.1021/bi301570m. Epub
            2013 Feb 15.
  PUBMED    23363317
REFERENCE   2  (bases 1 to 7803)
  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"; doi: "10.1021/bi301570m";
            journalName: "Biochemistry"; date: "2013-03-5- 5"; volume: "52"; 
            issue: "9"; pages: "1568-82"
COMMENT     Sequence Label: pETDuet-msGC-NT13
FEATURES             Location/Qualifiers
     source          1..7803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    3..27
                     /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             42..64
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             83..100
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     promoter        1393..1411
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1412..1436
                     /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)."
     CDS             2749..2793
                     /codon_start=1
                     /product="affinity and epitope tag derived from pancreatic 
                     ribonuclease A"
                     /label=S-Tag
                     /translation="KETAAAKFERQHMDS"
     primer_bind     complement(2831..2849)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2845..2892
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2929..3384
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3016..3035)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3226..3247
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(join(3499..4290,4291..4359))
                     /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     4122..4141
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4360..4452)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      4533..5121
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5022..5041
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5275..5292
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    5307..5449
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(5375..5394)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5535..5557
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5551..5742)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(6314..7396)
                     /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     7358..7377
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(7397..7474)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     7680..7699
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        join(7787..7803,1..2)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggggaattgt gagcggataa caattcccct ctagaaataa ttttgtttaa ctttaagaag
       61 gagatatacc atgggcagca gccatcacca tcatcaccac agccaggatc cgctcactct
      121 taagcatatg agtgaggctt tgcaactcct gactgctcca tcgaatgaat gcctccatgc
      181 agctgtgacg tcactcacga agaatcaatc agatcactac cataaataca attgtttgcg
      241 tagattacca gacgacgtaa aaacttgtag aaattacgcg tacctacaag aaatttacga
      301 cgccgtgcga gcaactgaca gtgtcaacac gaaggatttt atggcaaaat taggggaata
      361 tttaatttta acagccttct cgcacaattg tcggttagag agagcgttta aatgtttagg
      421 cacaaattta acagaatttc tgaccacact agacagtgtt catgacgtgt tgcacgatca
      481 ggacacaccg cttaaagatg aaacgatgga gtacgaagcg aatttcgtat gcacgacctc
      541 gcaagaaggg aagatacagt tacacttgac cacggaaagc gaaccagtgg cgtacctttt
      601 agtaggcagt ctgaaagcca tagcgaaacg actgtatgat acacagacag acataagact
      661 gaggagctat accaatgatc ctaggaggtt taggtacgaa ataaatgcag tcccactaca
      721 ccagaaatca aaagaagata gctgcgaact tgtcaacgag gcagcttctg ttgccacctc
      781 gaccaaagtg acagatttga agatcggcgt ggctagcttc tgcaaagcgt ttccatggca
      841 tttcatcact gataaaaggc tggaactcgt tcaattggga gctggattca tgcgcctctt
      901 cggaacccac ttggcaacac acgggtcttc gctgggcacg tacttcagac tcctgcgtcc
      961 aagaggagtg ccgctggact ttcgcgagat cctcaagcgt gtcaacacac cgttcatgtt
     1021 ttgcctaaag atgcctggtt cgacagcact tgccgagggt ttggagatca aaggacaaat
     1081 ggtgttctgt gccgagtcag attctttgct gtttgtcggg tccccgttct tggacggtct
     1141 cgaaggtctg actggaaggg gactcttcat atccgatata cctcttcatg atgcgacgag
     1201 agatgtcatc ttagtcggag aacaggctag ggctcaggat ggtttgcgaa gacgaatgga
     1261 caagctgaag aactcaatag aagaagcgag caaagcagta gacaaggaac gagagaagtg
     1321 aaagcttgcg gccgcataat gcttaagtcg aacagaaagt aatcgtattg tacacggccg
     1381 cataatcgaa attaatacga ctcactatag gggaattgtg agcggataac aattccccat
     1441 cttagtatat tagttaagta taagaaggag atatacatat gtacgggttt gtgaactatg
     1501 ccctcgagct ccttgtgatg aaaacttttg acgaagaaac atgggagacg ataaagaaaa
     1561 aggccgacgt ggctatggaa ggcagtttcc tagtgagaca gatttatgag gatgagatca
     1621 cttataattt gatcaccgct gccgttgagg tcttgcaaat acccgctgac gctatattgg
     1681 agctatttgg aaaaacgttc ttcgagtttt gccaagattc cgggtatgat aaaattctgc
     1741 aagtacttgg agccacgcca cgagattttc tacagaatct ggacggcctg cacgatcatc
     1801 tcgggacatt atatcctggc atgagatccc cctcgttccg atgcaccgag cgtcctgaag
     1861 atggcgccct tgtgcttcat tattactccg acaggcctgg tctggagcat atcgttattg
     1921 gaattgtaaa gactgtggcg agcaaactcc ataacaccga agtgaaggtg gaaatcctaa
     1981 agaccaaaga ggaatgtgac cacgtccagt tcttgatcac tgaaacttcc accaccggta
     2041 gggtgtctgc acctgagata gcagagattg aaacgttgtc tcttgaacca aaagtgagtc
     2101 cagcgacatt ctgccgagtg tttccattcc atttgatgtt tgatcgtgat ctgaacatcg
     2161 tccaagcggg acgtactgtg tctcgtctac tcccaagagt taccagacca ggctgtaaga
     2221 ttactgatgt cttggatact gtgcgtcccc acttagagat gacgtttgct aacgtcctgg
     2281 ctcatattaa cacggtctac gttttgaaaa ccaaacctga agagatgagt gtcactgatc
     2341 ctcatgagga aatcgcttct cttcgcttga aaggtcaaat gctgtacatt ccagaaacag
     2401 acgtggttgt attccagtgt tatccaagtg tgaccaattt ggacgatctg actcgccgcg
     2461 gcctatgcat tgctgacatt ccacttcacg atgcgacacg agacctggtt ctcatgtccg
     2521 agcaatttga agctgattat aagctgaccc agaatctgga agttttgact gacaaacttc
     2581 aacaaacatt ccgagagctg gaattggaaa aacagaagta ggacaggctt ctttactcag
     2641 tgctgcctat aagcgtggcc actgaacttc gtcaccgtcg ctgaaagggc gaattctgca
     2701 gattcggccg gccacgcgat cgctgacgtc ggtaccctcg agtctggtaa agaaaccgct
     2761 gctgcgaaat ttgaacgcca gcacatggac tcgtctacta gcgcagctta attaacctag
     2821 gctgctgcca ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg
     2881 aggggttttt tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta
     2941 gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca
     3001 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     3061 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     3121 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     3181 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     3241 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     3301 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     3361 acaaaatatt aacgtttaca atttctggcg gcacgatggc atgagattat caaaaaggat
     3421 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3481 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3541 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3601 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     3661 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     3721 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     3781 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     3841 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     3901 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     3961 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4021 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4081 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4141 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4201 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4261 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4321 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatcatg
     4381 attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa tgtatttaga
     4441 aaaataaaca aataggtcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg
     4501 tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc
     4561 tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag
     4621 ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc
     4681 cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac
     4741 ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc
     4801 gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt
     4861 tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt
     4921 gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc
     4981 ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt
     5041 tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca
     5101 ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt
     5161 tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt
     5221 attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag
     5281 tcagtgagcg aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc
     5341 ggtatttcac accgcatata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     5401 taagccagta tacactccgc tatcgctacg tgactgggtc atggctgcgc cccgacaccc
     5461 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5521 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5581 cgcgaggcag ctgcggtaaa gctcatcagc gtggtcgtga agcgattcac agatgtctgc
     5641 ctgttcatcc gcgtccagct cgttgagttt ctccagaagc gttaatgtct ggcttctgat
     5701 aaagcgggcc atgttaaggg cggttttttc ctgtttggtc actgatgcct ccgtgtaagg
     5761 gggatttctg ttcatggggg taatgatacc gatgaaacga gagaggatgc tcacgatacg
     5821 ggttactgat gatgaacatg cccggttact ggaacgttgt gagggtaaac aactggcggt
     5881 atggatgcgg cgggaccaga gaaaaatcac tcagggtcaa tgccagcgct tcgttaatac
     5941 agatgtaggt gttccacagg gtagccagca gcatcctgcg atgcagatcc ggaacataat
     6001 ggtgcagggc gctgacttcc gcgtttccag actttacgaa acacggaaac cgaagaccat
     6061 tcatgttgtt gctcaggtcg cagacgtttt gcagcagcag tcgcttcacg ttcgctcgcg
     6121 tatcggtgat tcattctgct aaccagtaag gcaaccccgc cagcctagcc gggtcctcaa
     6181 cgacaggagc acgatcatgc tagtcatgcc ccgcgcccac cggaaggagc tgactgggtt
     6241 gaaggctctc aagggcatcg gtcgagatcc cggtgcctaa tgagtgagct aacttacatt
     6301 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     6361 atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgccagg gtggtttttc
     6421 ttttcaccag tgagacgggc aacagctgat tgcccttcac cgcctggccc tgagagagtt
     6481 gcagcaagcg gtccacgctg gtttgcccca gcaggcgaaa atcctgtttg atggtggtta
     6541 acggcgggat ataacatgag ctgtcttcgg tatcgtcgta tcccactacc gagatgtccg
     6601 caccaacgcg cagcccggac tcggtaatgg cgcgcattgc gcccagcgcc atctgatcgt
     6661 tggcaaccag catcgcagtg ggaacgatgc cctcattcag catttgcatg gtttgttgaa
     6721 aaccggacat ggcactccag tcgccttccc gttccgctat cggctgaatt tgattgcgag
     6781 tgagatattt atgccagcca gccagacgca gacgcgccga gacagaactt aatgggcccg
     6841 ctaacagcgc gatttgctgg tgacccaatg cgaccagatg ctccacgccc agtcgcgtac
     6901 cgtcttcatg ggagaaaata atactgttga tgggtgtctg gtcagagaca tcaagaaata
     6961 acgccggaac attagtgcag gcagcttcca cagcaatggc atcctggtca tccagcggat
     7021 agttaatgat cagcccactg acgcgttgcg cgagaagatt gtgcaccgcc gctttacagg
     7081 cttcgacgcc gcttcgttct accatcgaca ccaccacgct ggcacccagt tgatcggcgc
     7141 gagatttaat cgccgcgaca atttgcgacg gcgcgtgcag ggccagactg gaggtggcaa
     7201 cgccaatcag caacgactgt ttgcccgcca gttgttgtgc cacgcggttg ggaatgtaat
     7261 tcagctccgc catcgccgct tccacttttt cccgcgtttt cgcagaaacg tggctggcct
     7321 ggttcaccac gcgggaaacg gtctgataag agacaccggc atactctgcg acatcgtata
     7381 acgttactgg tttcacattc accaccctga attgactctc ttccgggcgc tatcatgcca
     7441 taccgcgaaa ggttttgcgc cattcgatgg tgtccgggat ctcgacgctc tcccttatgc
     7501 gactcctgca ttaggaagca gcccagtagt aggttgaggc cgttgagcac cgccgccgca
     7561 aggaatggtg catgcaagga gatggcgccc aacagtcccc cggccacggg gcctgccacc
     7621 atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc ttccccatcg
     7681 gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat gccggccacg
     7741 atgcgtccgg cgtagaggat cgagatcgat ctcgatcccg cgaaattaat acgactcact
     7801 ata
//
