LOCUS       sequence-317611-        6324 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Hydrocarbon Structures for Fuels and Chemicals.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6324)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6324)
  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"
COMMENT     Sequence Label: JGI_017977
FEATURES             Location/Qualifiers
     source          1..6324
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    1..25
                     /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             40..62
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             81..98
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             108..125
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1170..1187
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1240..1258)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1254..1301
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1338..1793
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1425..1444)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1635..1656
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1885..2700)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1885..2700)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2608..2627
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2692..2711
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2822..3410
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3311..3330
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3564..3581
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3596..3735
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3661..3680)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3821..3843
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3837..4028)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4837..5919)
                     /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     5881..5900
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5920..5997)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6203..6222
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     join(6306..6324,1)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        6306..6324
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
       61 gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg
      121 gcagccatat gaagggctgc gcggtgtcaa ataactttca gtatcagttc aatgatgtag
      181 ccattgttag tatagaagca tgtgacgcgc cagtggtggt tactagtgct gccatcgacg
      241 aagccttggc gccatttctt gaacgcgttg gcgccgaaac tggtttactg cagaaattgg
      301 caggtattga agagaggcgc cagtggcccg acgatattag tttcattgaa gcgtctgcct
      361 tggcaggaga gaaggccatc gcggcaagtg gtattgaccc gagccgcatt ggcatgataa
      421 gcaatacgtc tgtttgccgg gatcgtctgg agccgtcttc ggctgttagc gtgcacgacc
      481 gccttggact ctcacctcac tgcattaact ttgacctgtc caacgcgtgt ctgggctttg
      541 tgaacgctat gcatcttgcc gggactctta ttgaatccgg gcagattgaa tatgcactga
      601 ttgtggacgg cgaaggtacg agggaaatcc aacagaatac cataaatcgg ctgcttgatg
      661 aacacagctc tatggcggat ctgttttcca attttgcgtc cctgacatta ggctctggga
      721 gcgcagccat gatcttaggt cgccattcaa ctaatccggg cagtcatcgt ctgggtcggg
      781 gcgagttccg agcagcaaca gaacatcatg atttatgcgt aggttccctc gaaagtatgc
      841 ggacggatac gcgtgcgttg cttgaagcgg ggacccagtt gggtaagata tgttgggacc
      901 aggccgactc tgatgactgg ggtgatatgg atagatacat tatacaccaa gtaagtcgtg
      961 ttcatactgc cgcgatggtg gagatcctgg ggattgacca caagagagtg ccgttgactt
     1021 tcccaaaata tggtaacata ggccctgcag cagttccatt taccctggca aaagaagctg
     1081 aaactttaaa tgaaggtgat cgggtactgt tagtcggcgt tggtagtggt ctgaatgctg
     1141 cttttctgga aattacgtgg tgactcgagc accaccacca ccaccactga gatccggctg
     1201 ctaacaaagc ccgaaaggaa gctgagttgg ctgctgccac cgctgagcaa taactagcat
     1261 aaccccttgg ggcctctaaa cgggtcttga ggggtttttt gctgaaagga ggaactatat
     1321 ccggattggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt
     1381 tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt tcgctttctt
     1441 cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc gggggctccc
     1501 tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg attagggtga
     1561 tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga cgttggagtc
     1621 cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc ctatctcggt
     1681 ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa aaaatgagct
     1741 gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa tttaggtggc
     1801 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     1861 atgtatccgc tcatgaatta attcttagaa aaactcatcg agcatcaaat gaaactgcaa
     1921 tttattcata tcaggattat caataccata tttttgaaaa agccgtttct gtaatgaagg
     1981 agaaaactca ccgaggcagt tccataggat ggcaagatcc tggtatcggt ctgcgattcc
     2041 gactcgtcca acatcaatac aacctattaa tttcccctcg tcaaaaataa ggttatcaag
     2101 tgagaaatca ccatgagtga cgactgaatc cggtgagaat ggcaaaagtt tatgcatttc
     2161 tttccagact tgttcaacag gccagccatt acgctcgtca tcaaaatcac tcgcatcaac
     2221 caaaccgtta ttcattcgtg attgcgcctg agcgagacga aatacgcgat cgctgttaaa
     2281 aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg aacactgcca gcgcatcaac
     2341 aatattttca cctgaatcag gatattcttc taatacctgg aatgctgttt tcccggggat
     2401 cgcagtggtg agtaaccatg catcatcagg agtacggata aaatgcttga tggtcggaag
     2461 aggcataaat tccgtcagcc agtttagtct gaccatctca tctgtaacat cattggcaac
     2521 gctacctttg ccatgtttca gaaacaactc tggcgcatcg ggcttcccat acaatcgata
     2581 gattgtcgca cctgattgcc cgacattatc gcgagcccat ttatacccat ataaatcagc
     2641 atccatgttg gaatttaatc gcggcctaga gcaagacgtt tcccgttgaa tatggctcat
     2701 aacacccctt gtattactgt ttatgtaagc agacagtttt attgttcatg accaaaatcc
     2761 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     2821 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     2881 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     2941 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     3001 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     3061 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     3121 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     3181 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     3241 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     3301 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     3361 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     3421 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     3481 cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc
     3541 gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga
     3601 tgcggtattt tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag
     3661 tacaatctgc tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac
     3721 tgggtcatgg ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt
     3781 ctgctcccgg catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag
     3841 aggttttcac cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg
     3901 tcgtgaagcg attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc
     3961 agaagcgtta atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt
     4021 ttggtcactg atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg
     4081 aaacgagaga ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa
     4141 cgttgtgagg gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag
     4201 ggtcaatgcc agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat
     4261 cctgcgatgc agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt
     4321 tacgaaacac ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag
     4381 cagcagtcgc ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa
     4441 ccccgccagc ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggggcc
     4501 gccatgccgg cgataatggc ctgcttctcg ccgaaacgtt tggtggcggg accagtgacg
     4561 aaggcttgag cgagggcgtg caagattccg aataccgcaa gcgacaggcc gatcatcgtc
     4621 gcgctccagc gaaagcggtc ctcgccgaaa atgacccaga gcgctgccgg cacctgtcct
     4681 acgagttgca tgataaagaa gacagtcata agtgcggcga cgatagtcat gccccgcgcc
     4741 caccggaagg agctgactgg gttgaaggct ctcaagggca tcggtcgaga tcccggtgcc
     4801 taatgagtga gctaacttac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga
     4861 aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     4921 attgggcgcc agggtggttt ttcttttcac cagtgagacg ggcaacagct gattgccctt
     4981 caccgcctgg ccctgagaga gttgcagcaa gcggtccacg ctggtttgcc ccagcaggcg
     5041 aaaatcctgt ttgatggtgg ttaacggcgg gatataacat gagctgtctt cggtatcgtc
     5101 gtatcccact accgagatat ccgcaccaac gcgcagcccg gactcggtaa tggcgcgcat
     5161 tgcgcccagc gccatctgat cgttggcaac cagcatcgca gtgggaacga tgccctcatt
     5221 cagcatttgc atggtttgtt gaaaaccgga catggcactc cagtcgcctt cccgttccgc
     5281 tatcggctga atttgattgc gagtgagata tttatgccag ccagccagac gcagacgcgc
     5341 cgagacagaa cttaatgggc ccgctaacag cgcgatttgc tggtgaccca atgcgaccag
     5401 atgctccacg cccagtcgcg taccgtcttc atgggagaaa ataatactgt tgatgggtgt
     5461 ctggtcagag acatcaagaa ataacgccgg aacattagtg caggcagctt ccacagcaat
     5521 ggcatcctgg tcatccagcg gatagttaat gatcagccca ctgacgcgtt gcgcgagaag
     5581 attgtgcacc gccgctttac aggcttcgac gccgcttcgt tctaccatcg acaccaccac
     5641 gctggcaccc agttgatcgg cgcgagattt aatcgccgcg acaatttgcg acggcgcgtg
     5701 cagggccaga ctggaggtgg caacgccaat cagcaacgac tgtttgcccg ccagttgttg
     5761 tgccacgcgg ttgggaatgt aattcagctc cgccatcgcc gcttccactt tttcccgcgt
     5821 tttcgcagaa acgtggctgg cctggttcac cacgcgggaa acggtctgat aagagacacc
     5881 ggcatactct gcgacatcgt ataacgttac tggtttcaca ttcaccaccc tgaattgact
     5941 ctcttccggg cgctatcatg ccataccgcg aaaggttttg cgccattcga tggtgtccgg
     6001 gatctcgacg ctctccctta tgcgactcct gcattaggaa gcagcccagt agtaggttga
     6061 ggccgttgag caccgccgcc gcaaggaatg gtgcatgcaa ggagatggcg cccaacagtc
     6121 ccccggccac ggggcctgcc accataccca cgccgaaaca agcgctcatg agcccgaagt
     6181 ggcgagcccg atcttcccca tcggtgatgt cggcgatata ggcgccagca accgcacctg
     6241 tggcgccggt gatgccggcc acgatgcgtc cggcgtagag gatcgagatc tcgatcccgc
     6301 gaaattaata cgactcacta tagg
//
