LOCUS       sequence-317198-        6321 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 6321)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6321)
  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_017973
FEATURES             Location/Qualifiers
     source          1..6321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    20..44
                     /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             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             1186..1203
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1256..1274)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1270..1317
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      1354..1809
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1441..1460)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1651..1672
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             complement(1901..2716)
                     /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(1901..2716)
                     /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     2624..2643
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2708..2727
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      2838..3426
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3327..3346
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3580..3597
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3612..3751
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3677..3696)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3837..3859
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3853..4044)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(4853..5935)
                     /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     5897..5916
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5936..6013)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6219..6238
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg gtcaaaacta tgaattatat tgctaaattg
      181 gttggtgttg gccaggcact gccagaacgc gtagttacat cagaagagat ggaggcaatg
      241 atgggctttg atcagttagg cataagaaaa ggaatggcca aaatactttc cggcgtttca
      301 gagcgtcgat tcgctagtga tgagattaat tcatctgatc tggccgtcga agctgcaaaa
      361 gaggcgatgg agatggcagg taagaaagcg gaagacattg acctggtcat cttttcggcc
      421 ttgacgcagg acgtagccga accggcaact gctaatatta tcatggacaa gctgaacatc
      481 cgtaatgcga agtcgtttga tattaagaac gcgtgcaatg cttttgtgtg cgctattgag
      541 attgcaaaca tgtatatcaa gtgcaatgat gtgaaaaatg ttcttattgc ttctggcgaa
      601 gttctgagta ggtttgtgaa aatggatctg ctgaaagacc cagccaaact gaaggatgtt
      661 aactcaacat ttagtgtggg tgatggcggc ggcgcgattg tactcgaacg tcaggaatct
      721 gaagaggatt ttggtattat tacagccttt cgtaccttcc cagatacttg gagtgacggg
      781 gtcctgtggg gaggtggtac catttaccca catgatccgg ataaatttta tttccagaat
      841 gaatcggaag aactaatgaa aattaacttt aaacgcgcta tgaaattctt tgaagaacag
      901 atggagaaaa tgaaaattgt cagggaggag atcgatctgt tcattccgac gcaaattaca
      961 aaatacatca taattaaatc tagcgaagtt ttgaaattgc cagatgatcg cgtggtcagc
     1021 cagataggca cccttgggaa catgtctact gcctcgattc cagtggccct cagccgcgca
     1081 gtaaacagcg ggcaactgaa gatcggcacc ggacagaaag tcgtattatt aggcgtcgcg
     1141 aatggtttta atgtgggttt ggtttatatg tccatctaac tcgagcacca ccaccaccac
     1201 cactgagatc cggctgctaa caaagcccga aaggaagctg agttggctgc tgccaccgct
     1261 gagcaataac tagcataacc ccttggggcc tctaaacggg tcttgagggg ttttttgctg
     1321 aaaggaggaa ctatatccgg attggcgaat gggacgcgcc ctgtagcggc gcattaagcg
     1381 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     1441 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     1501 taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     1561 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     1621 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     1681 tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt
     1741 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc
     1801 ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt
     1861 ctaaatacat tcaaatatgt atccgctcat gaattaattc ttagaaaaac tcatcgagca
     1921 tcaaatgaaa ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc
     1981 gtttctgtaa tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt
     2041 atcggtctgc gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa
     2101 aaataaggtt atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatggca
     2161 aaagtttatg catttctttc cagacttgtt caacaggcca gccattacgc tcgtcatcaa
     2221 aatcactcgc atcaaccaaa ccgttattca ttcgtgattg cgcctgagcg agacgaaata
     2281 cgcgatcgct gttaaaagga caattacaaa caggaatcga atgcaaccgg cgcaggaaca
     2341 ctgccagcgc atcaacaata ttttcacctg aatcaggata ttcttctaat acctggaatg
     2401 ctgttttccc ggggatcgca gtggtgagta accatgcatc atcaggagta cggataaaat
     2461 gcttgatggt cggaagaggc ataaattccg tcagccagtt tagtctgacc atctcatctg
     2521 taacatcatt ggcaacgcta cctttgccat gtttcagaaa caactctggc gcatcgggct
     2581 tcccatacaa tcgatagatt gtcgcacctg attgcccgac attatcgcga gcccatttat
     2641 acccatataa atcagcatcc atgttggaat ttaatcgcgg cctagagcaa gacgtttccc
     2701 gttgaatatg gctcataaca ccccttgtat tactgtttat gtaagcagac agttttattg
     2761 ttcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     2821 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     2881 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     2941 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg
     3001 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     3061 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     3121 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     3181 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     3241 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     3301 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     3361 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     3421 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     3481 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     3541 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     3601 gcggaagagc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc
     3661 aatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag tatacactcc
     3721 gctatcgcta cgtgactggg tcatggctgc gccccgacac ccgccaacac ccgctgacgc
     3781 gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg
     3841 gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgaggc agctgcggta
     3901 aagctcatca gcgtggtcgt gaagcgattc acagatgtct gcctgttcat ccgcgtccag
     3961 ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg ataaagcggg ccatgttaag
     4021 ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa gggggatttc tgttcatggg
     4081 ggtaatgata ccgatgaaac gagagaggat gctcacgata cgggttactg atgatgaaca
     4141 tgcccggtta ctggaacgtt gtgagggtaa acaactggcg gtatggatgc ggcgggacca
     4201 gagaaaaatc actcagggtc aatgccagcg cttcgttaat acagatgtag gtgttccaca
     4261 gggtagccag cagcatcctg cgatgcagat ccggaacata atggtgcagg gcgctgactt
     4321 ccgcgtttcc agactttacg aaacacggaa accgaagacc attcatgttg ttgctcaggt
     4381 cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg cgtatcggtg attcattctg
     4441 ctaaccagta aggcaacccc gccagcctag ccgggtcctc aacgacagga gcacgatcat
     4501 gcgcacccgt ggggccgcca tgccggcgat aatggcctgc ttctcgccga aacgtttggt
     4561 ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag attccgaata ccgcaagcga
     4621 caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg ccgaaaatga cccagagcgc
     4681 tgccggcacc tgtcctacga gttgcatgat aaagaagaca gtcataagtg cggcgacgat
     4741 agtcatgccc cgcgcccacc ggaaggagct gactgggttg aaggctctca agggcatcgg
     4801 tcgagatccc ggtgcctaat gagtgagcta acttacatta attgcgttgc gctcactgcc
     4861 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4921 gagaggcggt ttgcgtattg ggcgccaggg tggtttttct tttcaccagt gagacgggca
     4981 acagctgatt gcccttcacc gcctggccct gagagagttg cagcaagcgg tccacgctgg
     5041 tttgccccag caggcgaaaa tcctgtttga tggtggttaa cggcgggata taacatgagc
     5101 tgtcttcggt atcgtcgtat cccactaccg agatatccgc accaacgcgc agcccggact
     5161 cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt ggcaaccagc atcgcagtgg
     5221 gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa accggacatg gcactccagt
     5281 cgccttcccg ttccgctatc ggctgaattt gattgcgagt gagatattta tgccagccag
     5341 ccagacgcag acgcgccgag acagaactta atgggcccgc taacagcgcg atttgctggt
     5401 gacccaatgc gaccagatgc tccacgccca gtcgcgtacc gtcttcatgg gagaaaataa
     5461 tactgttgat gggtgtctgg tcagagacat caagaaataa cgccggaaca ttagtgcagg
     5521 cagcttccac agcaatggca tcctggtcat ccagcggata gttaatgatc agcccactga
     5581 cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc ttcgacgccg cttcgttcta
     5641 ccatcgacac caccacgctg gcacccagtt gatcggcgcg agatttaatc gccgcgacaa
     5701 tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac gccaatcagc aacgactgtt
     5761 tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt cagctccgcc atcgccgctt
     5821 ccactttttc ccgcgttttc gcagaaacgt ggctggcctg gttcaccacg cgggaaacgg
     5881 tctgataaga gacaccggca tactctgcga catcgtataa cgttactggt ttcacattca
     5941 ccaccctgaa ttgactctct tccgggcgct atcatgccat accgcgaaag gttttgcgcc
     6001 attcgatggt gtccgggatc tcgacgctct cccttatgcg actcctgcat taggaagcag
     6061 cccagtagta ggttgaggcc gttgagcacc gccgccgcaa ggaatggtgc atgcaaggag
     6121 atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc gaaacaagcg
     6181 ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc gatataggcg
     6241 ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc gtagaggatc
     6301 gagatctcga tcccgcgaaa t
//
