LOCUS       sequence-317192-        6312 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 6312)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6312)
  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_018089
FEATURES             Location/Qualifiers
     source          1..6312
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    132..271
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(197..216)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     357..379
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(373..564)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(1373..2455)
                     /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     2417..2436
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(2456..2533)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     2739..2758
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     2842..2861
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2842..2860
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2861..2885
                     /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             2900..2922
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             2941..2958
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2968..2985
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             4018..4035
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4088..4106)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      4102..4149
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      4186..4641
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4273..4292)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4483..4504
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(4733..5548)
                     /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(4733..5548)
                     /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     5456..5475
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5540..5559
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5670..6258
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6159..6178
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
       61 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      121 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
      181 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
      241 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
      301 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
      361 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
      421 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
      481 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
      541 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
      601 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
      661 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
      721 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
      781 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
      841 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
      901 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
      961 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     1021 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     1081 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     1141 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     1201 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     1261 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     1321 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     1381 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     1441 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     1501 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     1561 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     1621 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     1681 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     1741 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     1801 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     1861 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     1921 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     1981 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     2041 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     2101 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     2161 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     2221 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     2281 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     2341 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     2401 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     2461 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     2521 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     2581 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     2641 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     2701 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     2761 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     2821 gagatctcga tcccgcgaaa ttaatacgac tcactatagg ggaattgtga gcggataaca
     2881 attcccctct agaaataatt ttgtttaact ttaagaagga gatataccat gggcagcagc
     2941 catcatcatc atcatcacag cagcggcctg gtgccgcgcg gcagccatat gagtgaaaat
     3001 gcgacttatc ggttggcaaa tactacgatt ctgtccgttt cagcactgga agccccacag
     3061 gtggttagct ccgcagaact ggatgagtcg ttagctccag cgtacgcgcg ccatggcgtt
     3121 aaagcaggtc agatcgaacg tttggcgggg gtagttcaga gacggtggta ccccccggag
     3181 gctgattata cagatggggc tgttgatgtt ggacgtgccg ctatggccga tgcgggtgtt
     3241 gaagcaggcg aaattggcct ggtaataaac gcgtcggtca ctcgcccgta tcttgaaccg
     3301 ggtattagcg cccgcgttca tcacgaattg ggcctgcccc gcagagcaat tgcatttgac
     3361 ctcaccaatg cgtgcctcgg agtagctaac gcaatgcatg tggcggggac tatgatagac
     3421 gctggacaga tcgacaatgc gctgattgtt gcctctgaag gttctcgtca aatgcaggaa
     3481 tcctctatca aatatcttct ttccgatgat agcacacgtg aggatgttaa agaggcgttt
     3541 gcaaccatga cattaggctc aggtgctgtc ggtctggtgc tgggcagatc cgatgcccga
     3601 ccggacggcc atcgcgtgat agggggtgtg actcgtgccg gctcggaaca ccataccctg
     3661 tgtgtgggcg gaatggatgg tatgcgcacg gatagctctc gcctattcag agaaggctta
     3721 acacttgcaa ccgaggcatg gaaggacgca cgcggcgact tcgattggga tgatatggac
     3781 tggtacgttg ctcatcaaac ctctaccatt catatagaag cactttgcaa aaccttggga
     3841 ctgccgcaag aacgattccc gatgaccttg ccgacgtatg gtaatattgg tccggtcgcg
     3901 atgccgtata ctttagcctt ccatcaagat aatatagaac gggggcagcg cgttttgtta
     3961 atggggattg gttcaggcct gaacacgagt atgacagagg tagcctggtg actcgagcac
     4021 caccaccacc accactgaga tccggctgct aacaaagccc gaaaggaagc tgagttggct
     4081 gctgccaccg ctgagcaata actagcataa ccccttgggg cctctaaacg ggtcttgagg
     4141 ggttttttgc tgaaaggagg aactatatcc ggattggcga atgggacgcg ccctgtagcg
     4201 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     4261 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     4321 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
     4381 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     4441 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     4501 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
     4561 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattttaaca
     4621 aaatattaac gcttacaatt taggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4681 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgaattaat tcttagaaaa
     4741 actcatcgag catcaaatga aactgcaatt tattcatatc aggattatca ataccatatt
     4801 tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc gaggcagttc cataggatgg
     4861 caagatcctg gtatcggtct gcgattccga ctcgtccaac atcaatacaa cctattaatt
     4921 tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc atgagtgacg actgaatccg
     4981 gtgagaatgg caaaagttta tgcatttctt tccagacttg ttcaacaggc cagccattac
     5041 gctcgtcatc aaaatcactc gcatcaacca aaccgttatt cattcgtgat tgcgcctgag
     5101 cgagacgaaa tacgcgatcg ctgttaaaag gacaattaca aacaggaatc gaatgcaacc
     5161 ggcgcaggaa cactgccagc gcatcaacaa tattttcacc tgaatcagga tattcttcta
     5221 atacctggaa tgctgttttc ccggggatcg cagtggtgag taaccatgca tcatcaggag
     5281 tacggataaa atgcttgatg gtcggaagag gcataaattc cgtcagccag tttagtctga
     5341 ccatctcatc tgtaacatca ttggcaacgc tacctttgcc atgtttcaga aacaactctg
     5401 gcgcatcggg cttcccatac aatcgataga ttgtcgcacc tgattgcccg acattatcgc
     5461 gagcccattt atacccatat aaatcagcat ccatgttgga atttaatcgc ggcctagagc
     5521 aagacgtttc ccgttgaata tggctcataa caccccttgt attactgttt atgtaagcag
     5581 acagttttat tgttcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5641 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     5701 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     5761 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgtcctt
     5821 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     5881 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     5941 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6001 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6061 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6121 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6181 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6241 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6301 tggccttttg ct
//
