LOCUS       sequence-316780-        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_018121
FEATURES             Location/Qualifiers
     source          1..6321
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..816)
                     /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(1..816)
                     /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     724..743
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     808..827
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      938..1526
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1427..1446
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1680..1697
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    1712..1851
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(1777..1796)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     1937..1959
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(1953..2144)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(2953..4035)
                     /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     3997..4016
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(4036..4113)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     4319..4338
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     primer_bind     4422..4441
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4422..4440
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    4441..4465
                     /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             4480..4502
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             4521..4538
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             4548..4565
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             5607..5624
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5677..5695)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      5691..5738
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      5775..6230
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5862..5881)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6072..6093
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 ttagaaaaac tcatcgagca tcaaatgaaa ctgcaattta ttcatatcag gattatcaat
       61 accatatttt tgaaaaagcc gtttctgtaa tgaaggagaa aactcaccga ggcagttcca
      121 taggatggca agatcctggt atcggtctgc gattccgact cgtccaacat caatacaacc
      181 tattaatttc ccctcgtcaa aaataaggtt atcaagtgag aaatcaccat gagtgacgac
      241 tgaatccggt gagaatggca aaagtttatg catttctttc cagacttgtt caacaggcca
      301 gccattacgc tcgtcatcaa aatcactcgc atcaaccaaa ccgttattca ttcgtgattg
      361 cgcctgagcg agacgaaata cgcgatcgct gttaaaagga caattacaaa caggaatcga
      421 atgcaaccgg cgcaggaaca ctgccagcgc atcaacaata ttttcacctg aatcaggata
      481 ttcttctaat acctggaatg ctgttttccc ggggatcgca gtggtgagta accatgcatc
      541 atcaggagta cggataaaat gcttgatggt cggaagaggc ataaattccg tcagccagtt
      601 tagtctgacc atctcatctg taacatcatt ggcaacgcta cctttgccat gtttcagaaa
      661 caactctggc gcatcgggct tcccatacaa tcgatagatt gtcgcacctg attgcccgac
      721 attatcgcga gcccatttat acccatataa atcagcatcc atgttggaat ttaatcgcgg
      781 cctagagcaa gacgtttccc gttgaatatg gctcataaca ccccttgtat tactgtttat
      841 gtaagcagac agttttattg ttcatgacca aaatccctta acgtgagttt tcgttccact
      901 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
      961 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1021 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1081 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1141 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1201 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1261 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1321 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1381 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1441 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1501 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1561 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1621 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1681 gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc cttacgcatc
     1741 tgtgcggtat ttcacaccgc aatggtgcac tctcagtaca atctgctctg atgccgcata
     1801 gttaagccag tatacactcc gctatcgcta cgtgactggg tcatggctgc gccccgacac
     1861 ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga
     1921 caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa
     1981 cgcgcgaggc agctgcggta aagctcatca gcgtggtcgt gaagcgattc acagatgtct
     2041 gcctgttcat ccgcgtccag ctcgttgagt ttctccagaa gcgttaatgt ctggcttctg
     2101 ataaagcggg ccatgttaag ggcggttttt tcctgtttgg tcactgatgc ctccgtgtaa
     2161 gggggatttc tgttcatggg ggtaatgata ccgatgaaac gagagaggat gctcacgata
     2221 cgggttactg atgatgaaca tgcccggtta ctggaacgtt gtgagggtaa acaactggcg
     2281 gtatggatgc ggcgggacca gagaaaaatc actcagggtc aatgccagcg cttcgttaat
     2341 acagatgtag gtgttccaca gggtagccag cagcatcctg cgatgcagat ccggaacata
     2401 atggtgcagg gcgctgactt ccgcgtttcc agactttacg aaacacggaa accgaagacc
     2461 attcatgttg ttgctcaggt cgcagacgtt ttgcagcagc agtcgcttca cgttcgctcg
     2521 cgtatcggtg attcattctg ctaaccagta aggcaacccc gccagcctag ccgggtcctc
     2581 aacgacagga gcacgatcat gcgcacccgt ggggccgcca tgccggcgat aatggcctgc
     2641 ttctcgccga aacgtttggt ggcgggacca gtgacgaagg cttgagcgag ggcgtgcaag
     2701 attccgaata ccgcaagcga caggccgatc atcgtcgcgc tccagcgaaa gcggtcctcg
     2761 ccgaaaatga cccagagcgc tgccggcacc tgtcctacga gttgcatgat aaagaagaca
     2821 gtcataagtg cggcgacgat agtcatgccc cgcgcccacc ggaaggagct gactgggttg
     2881 aaggctctca agggcatcgg tcgagatccc ggtgcctaat gagtgagcta acttacatta
     2941 attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa
     3001 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgccaggg tggtttttct
     3061 tttcaccagt gagacgggca acagctgatt gcccttcacc gcctggccct gagagagttg
     3121 cagcaagcgg tccacgctgg tttgccccag caggcgaaaa tcctgtttga tggtggttaa
     3181 cggcgggata taacatgagc tgtcttcggt atcgtcgtat cccactaccg agatatccgc
     3241 accaacgcgc agcccggact cggtaatggc gcgcattgcg cccagcgcca tctgatcgtt
     3301 ggcaaccagc atcgcagtgg gaacgatgcc ctcattcagc atttgcatgg tttgttgaaa
     3361 accggacatg gcactccagt cgccttcccg ttccgctatc ggctgaattt gattgcgagt
     3421 gagatattta tgccagccag ccagacgcag acgcgccgag acagaactta atgggcccgc
     3481 taacagcgcg atttgctggt gacccaatgc gaccagatgc tccacgccca gtcgcgtacc
     3541 gtcttcatgg gagaaaataa tactgttgat gggtgtctgg tcagagacat caagaaataa
     3601 cgccggaaca ttagtgcagg cagcttccac agcaatggca tcctggtcat ccagcggata
     3661 gttaatgatc agcccactga cgcgttgcgc gagaagattg tgcaccgccg ctttacaggc
     3721 ttcgacgccg cttcgttcta ccatcgacac caccacgctg gcacccagtt gatcggcgcg
     3781 agatttaatc gccgcgacaa tttgcgacgg cgcgtgcagg gccagactgg aggtggcaac
     3841 gccaatcagc aacgactgtt tgcccgccag ttgttgtgcc acgcggttgg gaatgtaatt
     3901 cagctccgcc atcgccgctt ccactttttc ccgcgttttc gcagaaacgt ggctggcctg
     3961 gttcaccacg cgggaaacgg tctgataaga gacaccggca tactctgcga catcgtataa
     4021 cgttactggt ttcacattca ccaccctgaa ttgactctct tccgggcgct atcatgccat
     4081 accgcgaaag gttttgcgcc attcgatggt gtccgggatc tcgacgctct cccttatgcg
     4141 actcctgcat taggaagcag cccagtagta ggttgaggcc gttgagcacc gccgccgcaa
     4201 ggaatggtgc atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca
     4261 tacccacgcc gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg
     4321 tgatgtcggc gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga
     4381 tgcgtccggc gtagaggatc gagatctcga tcccgcgaaa ttaatacgac tcactatagg
     4441 ggaattgtga gcggataaca attcccctct agaaataatt ttgtttaact ttaagaagga
     4501 gatataccat gggcagcagc catcatcatc atcatcacag cagcggcctg gtgccgcgcg
     4561 gcagccatat gctgctaaac ggcaatgcaa cgtttcgcca tcgaaatact tcgttattag
     4621 gcctggcctc cgttttggcc ccgcatactg ttacctctgt agagattgac gatcgtttaa
     4681 aaccggtatt aagccgtctg cgtctgccaa cgggacttct ccaaagagtt gccggtgtgc
     4741 tggagcggcg caactgggat acatcaatga gtttcgatgc ggcggccaca gaagcaggtc
     4801 gcacagcact ggcacaggca ggagtgcagc cctcccagat aggtcttctg atcaatacgt
     4861 ccgtcactcg cgcgcatttg gagcctagcg tcgccgtgag tattcataac ggtctgggcc
     4921 tgccgtcctc cgcactgaat tttgacatag ccaatgcatg tctgggcttt gtgaatgcga
     4981 tgaccttagc aggccatctg attgatagcg gtcagattga ctacgcaatg atagtcgatg
     5041 gtgaagatgc cggggagatc cgccataaca ccgtcgcgcg tctgttacgt ccagagacta
     5101 cccgtgctga tttcctgagc gaattcccat ctctgacttt aggtgccgga gccgcagctg
     5161 ccgttctggg tcggacatcg gatcatccgg aaggacatcg tattcttgga ggagttacac
     5221 gtgcagcgac ccagcaccat gaactctgca ttggcgacgt cgatgggatg ttcaccgata
     5281 caaaggaact gcttcgtggc ggaatggaat tggttgtgga tgcatggaaa gaagcggccc
     5341 aagatgactg ggaatggtct gacatggacc gctatatttt gcatcaggtg agcgacgttc
     5401 atactaacgc cattgtcaag gctgcaaagt tagataagag cagagtccca ctgacttatc
     5461 cccggtatgg aaatgtcggc cctgctagta tacctattac cctggctgat caggcggata
     5521 gtttaagccg tggtgaccga gtactttgta tgggcgtagg cagcggatta aatacagcga
     5581 tgaccgagat tctctggtaa ctcgagcacc accaccacca ccactgagat ccggctgcta
     5641 acaaagcccg aaaggaagct gagttggctg ctgccaccgc tgagcaataa ctagcataac
     5701 cccttggggc ctctaaacgg gtcttgaggg gttttttgct gaaaggagga actatatccg
     5761 gattggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac
     5821 gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc
     5881 ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt
     5941 agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt agggtgatgg
     6001 ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac
     6061 gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta tctcggtcta
     6121 ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat
     6181 ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt aggtggcact
     6241 tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg
     6301 tatccgctca tgaattaatt c
//
