LOCUS       sequence-315448-        6306 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 6306)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6306)
  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_018073
FEATURES             Location/Qualifiers
     source          1..6306
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     796..815
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(835..912)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     1118..1137
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     1221..1240
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1221..1239
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1240..1264
                     /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             1279..1301
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1320..1337
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             1347..1364
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             2391..2408
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(2461..2479)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      2475..2522
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      2559..3014
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2646..2665)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2856..2877
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(3106..3921)
                     /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(3106..3921)
                     /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     3829..3848
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3913..3932
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4043..4631
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4532..4551
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4785..4802
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4817..4956
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4882..4901)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5042..5064
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(5058..5249)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(6058..6306,1..834))
                     /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"
ORIGIN
        1 gtcttcggta tcgtcgtatc ccactaccga gatatccgca ccaacgcgca gcccggactc
       61 ggtaatggcg cgcattgcgc ccagcgccat ctgatcgttg gcaaccagca tcgcagtggg
      121 aacgatgccc tcattcagca tttgcatggt ttgttgaaaa ccggacatgg cactccagtc
      181 gccttcccgt tccgctatcg gctgaatttg attgcgagtg agatatttat gccagccagc
      241 cagacgcaga cgcgccgaga cagaacttaa tgggcccgct aacagcgcga tttgctggtg
      301 acccaatgcg accagatgct ccacgcccag tcgcgtaccg tcttcatggg agaaaataat
      361 actgttgatg ggtgtctggt cagagacatc aagaaataac gccggaacat tagtgcaggc
      421 agcttccaca gcaatggcat cctggtcatc cagcggatag ttaatgatca gcccactgac
      481 gcgttgcgcg agaagattgt gcaccgccgc tttacaggct tcgacgccgc ttcgttctac
      541 catcgacacc accacgctgg cacccagttg atcggcgcga gatttaatcg ccgcgacaat
      601 ttgcgacggc gcgtgcaggg ccagactgga ggtggcaacg ccaatcagca acgactgttt
      661 gcccgccagt tgttgtgcca cgcggttggg aatgtaattc agctccgcca tcgccgcttc
      721 cactttttcc cgcgttttcg cagaaacgtg gctggcctgg ttcaccacgc gggaaacggt
      781 ctgataagag acaccggcat actctgcgac atcgtataac gttactggtt tcacattcac
      841 caccctgaat tgactctctt ccgggcgcta tcatgccata ccgcgaaagg ttttgcgcca
      901 ttcgatggtg tccgggatct cgacgctctc ccttatgcga ctcctgcatt aggaagcagc
      961 ccagtagtag gttgaggccg ttgagcaccg ccgccgcaag gaatggtgca tgcaaggaga
     1021 tggcgcccaa cagtcccccg gccacggggc ctgccaccat acccacgccg aaacaagcgc
     1081 tcatgagccc gaagtggcga gcccgatctt ccccatcggt gatgtcggcg atataggcgc
     1141 cagcaaccgc acctgtggcg ccggtgatgc cggccacgat gcgtccggcg tagaggatcg
     1201 agatctcgat cccgcgaaat taatacgact cactataggg gaattgtgag cggataacaa
     1261 ttcccctcta gaaataattt tgtttaactt taagaaggag atataccatg ggcagcagcc
     1321 atcatcatca tcatcacagc agcggcctgg tgccgcgcgg cagccatatg ctgtttaccc
     1381 atgttgctat agccggtctg gcccacatag acgcgccgca tgcattgtca tccgccgaca
     1441 tcaacgcgcg tttacagcct acaatggatc gcttgggaat taaaaccgat gtcctgcagg
     1501 atattgcagg aatatttagc agacggatgt gggatgatga aactcaaagt tcggacgccg
     1561 ccacgatggc ggcagagaaa gccttgctgg atgccaacgt cccgagggat aagatcggac
     1621 tgctggtgaa cacctccgtc tcacgggatt ttttggagcc gtcaactgca tccattgtta
     1681 gcggcaatct cggcctaggc gagaactgtc aaaattttga tgtagcaaac gcctgtctag
     1741 catttctgaa tggtatggac atcgcgtccc gcatgataga gcggggagac atcgaatatg
     1801 ctcttgtggt ggacggtgaa acagcgaact tggtctatga aaagaccctg actagactgt
     1861 cagatccgaa tgtgaccgag gagcagttcc gcaatgagct tgcgtctctc accttaggat
     1921 gtggtgccgc cgcaatggtg ttagccaggg cagagctggt cccggacgcc cctcgttaca
     1981 aagggggtgt gaccaaagcc gcgacgcaat ggaacactct gtgtcgaggt aatctagatc
     2041 ggatggtaac ggatacccga atgttactga ttgagggact taaattggcc cagaaaacgt
     2101 ttgttgcggc acgtctggcc ctgggctggg cggtcgagga aatggaccaa ttcgttatcc
     2161 accaagttag tcgtgtgcac acagaagctt ttatcaaagc gttcggaata gatccggcaa
     2221 aagtgcttac aatatttaat gaacatggaa atattggacc ggcatctgtg ccgatcgtgc
     2281 tgtccaaact caaagagatg ggtcgcctga aaaaagggaa tcggattgca ttgttaggaa
     2341 ttggatcggg cttaaactgc tcaatggcag aagtagtgtg gtaactcgag caccaccacc
     2401 accaccactg agatccggct gctaacaaag cccgaaagga agctgagttg gctgctgcca
     2461 ccgctgagca ataactagca taaccccttg gggcctctaa acgggtcttg aggggttttt
     2521 tgctgaaagg aggaactata tccggattgg cgaatgggac gcgccctgta gcggcgcatt
     2581 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     2641 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     2701 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     2761 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt
     2821 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac
     2881 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc
     2941 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta acaaaatatt
     3001 aacgcttaca atttaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     3061 tttttctaaa tacattcaaa tatgtatccg ctcatgaatt aattcttaga aaaactcatc
     3121 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     3181 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     3241 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     3301 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     3361 tggcaaaagt ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     3421 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcgagacg
     3481 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     3541 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     3601 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     3661 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     3721 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     3781 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     3841 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctag agcaagacgt
     3901 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     3961 tattgttcat gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg
     4021 tagaaaagat caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc
     4081 aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc
     4141 tttttccgaa ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt
     4201 agccgtagtt aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc
     4261 taatcctgtt accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact
     4321 caagacgata gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac
     4381 agcccagctt ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag
     4441 aaagcgccac gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg
     4501 gaacaggaga gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg
     4561 tcgggtttcg ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga
     4621 gcctatggaa aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt
     4681 ttgctcacat gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct
     4741 ttgagtgagc tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg
     4801 aggaagcgga agagcgcctg atgcggtatt ttctccttac gcatctgtgc ggtatttcac
     4861 accgcaatgg tgcactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatac
     4921 actccgctat cgctacgtga ctgggtcatg gctgcgcccc gacacccgcc aacacccgct
     4981 gacgcgccct gacgggcttg tctgctcccg gcatccgctt acagacaagc tgtgaccgtc
     5041 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gaggcagctg
     5101 cggtaaagct catcagcgtg gtcgtgaagc gattcacaga tgtctgcctg ttcatccgcg
     5161 tccagctcgt tgagtttctc cagaagcgtt aatgtctggc ttctgataaa gcgggccatg
     5221 ttaagggcgg ttttttcctg tttggtcact gatgcctccg tgtaaggggg atttctgttc
     5281 atgggggtaa tgataccgat gaaacgagag aggatgctca cgatacgggt tactgatgat
     5341 gaacatgccc ggttactgga acgttgtgag ggtaaacaac tggcggtatg gatgcggcgg
     5401 gaccagagaa aaatcactca gggtcaatgc cagcgcttcg ttaatacaga tgtaggtgtt
     5461 ccacagggta gccagcagca tcctgcgatg cagatccgga acataatggt gcagggcgct
     5521 gacttccgcg tttccagact ttacgaaaca cggaaaccga agaccattca tgttgttgct
     5581 caggtcgcag acgttttgca gcagcagtcg cttcacgttc gctcgcgtat cggtgattca
     5641 ttctgctaac cagtaaggca accccgccag cctagccggg tcctcaacga caggagcacg
     5701 atcatgcgca cccgtggggc cgccatgccg gcgataatgg cctgcttctc gccgaaacgt
     5761 ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc gaataccgca
     5821 agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa aatgacccag
     5881 agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat aagtgcggcg
     5941 acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc tctcaagggc
     6001 atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc gttgcgctca
     6061 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc
     6121 gcggggagag gcggtttgcg tattgggcgc cagggtggtt tttcttttca ccagtgagac
     6181 gggcaacagc tgattgccct tcaccgcctg gccctgagag agttgcagca agcggtccac
     6241 gctggtttgc cccagcaggc gaaaatcctg tttgatggtg gttaacggcg ggatataaca
     6301 tgagct
//
