LOCUS       sequence-315981-        6309 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 6309)
  TITLE     Hydrocarbon Structures for Fuels and Chemicals
REFERENCE   2  (bases 1 to 6309)
  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_018065
FEATURES             Location/Qualifiers
     source          1..6309
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             403..594
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(588..610)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    696..835
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     751..770
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(850..867)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1021..1609)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1101..1120)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(1720..1739)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             1731..2546
                     /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             1731..2546
                     /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     complement(1804..1823)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(2638..3093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2775..2796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2987..3006
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     terminator      complement(3130..3177)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     3173..3191
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(3244..3261)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(4291..4308)
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     CDS             complement(4318..4335)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             4354..4376
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4391..4415
                     /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)."
     primer_bind     complement(4415..4434)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4416..4434)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4518..4537)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        4743..4820
                     /gene="lacI"
                     /label=lacI promoter
     CDS             4821..5903
                     /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     complement(4840..4859)
                     /label=LacI-R
                     /note="LacI, reverse primer"
ORIGIN
        1 ggttagcaga atgaatcacc gatacgcgag cgaacgtgaa gcgactgctg ctgcaaaacg
       61 tctgcgacct gagcaacaac atgaatggtc ttcggtttcc gtgtttcgta aagtctggaa
      121 acgcggaagt cagcgccctg caccattatg ttccggatct gcatcgcagg atgctgctgg
      181 ctaccctgtg gaacacctac atctgtatta acgaagcgct ggcattgacc ctgagtgatt
      241 tttctctggt cccgccgcat ccataccgcc agttgtttac cctcacaacg ttccagtaac
      301 cgggcatgtt catcatcagt aacccgtatc gtgagcatcc tctctcgttt catcggtatc
      361 attaccccca tgaacagaaa tcccccttac acggaggcat cagtgaccaa acaggaaaaa
      421 accgccctta acatggcccg ctttatcaga agccagacat taacgcttct ggagaaactc
      481 aacgagctgg acgcggatga acaggcagac atctgtgaat cgcttcacga ccacgctgat
      541 gagctttacc gcagctgcct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
      601 cagctcccgg agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt
      661 cagggcgcgt cagcgggtgt tggcgggtgt cggggcgcag ccatgaccca gtcacgtagc
      721 gatagcggag tgtatactgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      781 accattgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca tcaggcgctc
      841 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
      901 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
      961 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     1021 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     1081 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     1141 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     1201 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     1261 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     1321 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     1381 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     1441 taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac
     1501 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     1561 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     1621 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     1681 catgaacaat aaaactgtct gcttacataa acagtaatac aaggggtgtt atgagccata
     1741 ttcaacggga aacgtcttgc tctaggccgc gattaaattc caacatggat gctgatttat
     1801 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgattgt
     1861 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     1921 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     1981 tcaagcattt tatccgtact cctgatgatg catggttact caccactgcg atccccggga
     2041 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     2101 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     2161 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     2221 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     2281 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     2341 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     2401 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     2461 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     2521 atttgatgct cgatgagttt ttctaagaat taattcatga gcggatacat atttgaatgt
     2581 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctaaa
     2641 ttgtaagcgt taatattttg ttaaaattcg cgttaaattt ttgttaaatc agctcatttt
     2701 ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc aaaagaatag accgagatag
     2761 ggttgagtgt tgttccagtt tggaacaaga gtccactatt aaagaacgtg gactccaacg
     2821 tcaaagggcg aaaaaccgtc tatcagggcg atggcccact acgtgaacca tcaccctaat
     2881 caagtttttt ggggtcgagg tgccgtaaag cactaaatcg gaaccctaaa gggagccccc
     2941 gatttagagc ttgacgggga aagccggcga acgtggcgag aaaggaaggg aagaaagcga
     3001 aaggagcggg cgctagggcg ctggcaagtg tagcggtcac gctgcgcgta accaccacac
     3061 ccgccgcgct taatgcgccg ctacagggcg cgtcccattc gccaatccgg atatagttcc
     3121 tcctttcagc aaaaaacccc tcaagacccg tttagaggcc ccaaggggtt atgctagtta
     3181 ttgctcagcg gtggcagcag ccaactcagc ttcctttcgg gctttgttag cagccggatc
     3241 tcagtggtgg tggtggtggt gctcgagtta ccaaggcggt acgtctaaaa tttcaaccac
     3301 tgctgcggac agggagaaac cggcaccagc cgccacgagc agtacaaggt ctcctggacc
     3361 gaccgcacgt tccttcacaa gatgctccag gccgagaaac tggtctccgg cgcctatatg
     3421 acccactaag cgtccgagat cccatgtgct ttgtttctcg gtgaagccca gtacatcgta
     3481 attctctggc cacacgcccc ggtggataag tggggttacc acgcggctga tatcctccac
     3541 actgacctgt gcttccgcta gtgccctccg ggtggtatcc ataagtgcag cttcaattct
     3601 ctcccagctc ccttctgctt caggtgtcgc cgcatgttgc agaccacgtt cacgcagccg
     3661 aagcggcatt tccagccccg gcgccgtcgc aaagggggtt gtacctcgat tccaacgttc
     3721 aagactatta tctgcgccca gggtggcgga caccacacgg gcgaacccct tgcgacctga
     3781 taagaccaag gcggtacccc catcggcaaa aaccaagttg agctgtacat tccagcggtc
     3841 gacccacggc ggcgcaaaat tatccccggt ggttaagagc acggcggagg taaaaccggc
     3901 atcaatatac ccagcagcga gacagaggga acccaatcca gcattgcatc tttgctgcag
     3961 atcaaagccg ggggcagccc tcccgacagt ttcgtttgca acatacgcgg cggcgggcca
     4021 catctcgtgg ccctgaaaac cacaagaaga atgcagcacc aggccaactt cgtcgcgctc
     4081 aatgcctact gcacgttcaa gagccagacg acctgcacgt actgccattt cagggcccgc
     4141 gactccatct gcgactgcta tagattcgta acccaggttt cgatgctcgt catcaactag
     4201 acctgcagct atcgcctcct ccatgggcac taaatctggt aaccaagaag ctgtactggc
     4261 gatatataaa tcgttctcaa agcgcatatg gctgccgcgc ggcaccaggc cgctgctgtg
     4321 atgatgatga tgatggctgc tgcccatggt atatctcctt cttaaagtta aacaaaatta
     4381 tttctagagg ggaattgtta tccgctcaca attcccctat agtgagtcgt attaatttcg
     4441 cgggatcgag atctcgatcc tctacgccgg acgcatcgtg gccggcatca ccggcgccac
     4501 aggtgcggtt gctggcgcct atatcgccga catcaccgat ggggaagatc gggctcgcca
     4561 cttcgggctc atgagcgctt gtttcggcgt gggtatggtg gcaggccccg tggccggggg
     4621 actgttgggc gccatctcct tgcatgcacc attccttgcg gcggcggtgc tcaacggcct
     4681 caacctacta ctgggctgct tcctaatgca ggagtcgcat aagggagagc gtcgagatcc
     4741 cggacaccat cgaatggcgc aaaacctttc gcggtatggc atgatagcgc ccggaagaga
     4801 gtcaattcag ggtggtgaat gtgaaaccag taacgttata cgatgtcgca gagtatgccg
     4861 gtgtctctta tcagaccgtt tcccgcgtgg tgaaccaggc cagccacgtt tctgcgaaaa
     4921 cgcgggaaaa agtggaagcg gcgatggcgg agctgaatta cattcccaac cgcgtggcac
     4981 aacaactggc gggcaaacag tcgttgctga ttggcgttgc cacctccagt ctggccctgc
     5041 acgcgccgtc gcaaattgtc gcggcgatta aatctcgcgc cgatcaactg ggtgccagcg
     5101 tggtggtgtc gatggtagaa cgaagcggcg tcgaagcctg taaagcggcg gtgcacaatc
     5161 ttctcgcgca acgcgtcagt gggctgatca ttaactatcc gctggatgac caggatgcca
     5221 ttgctgtgga agctgcctgc actaatgttc cggcgttatt tcttgatgtc tctgaccaga
     5281 cacccatcaa cagtattatt ttctcccatg aagacggtac gcgactgggc gtggagcatc
     5341 tggtcgcatt gggtcaccag caaatcgcgc tgttagcggg cccattaagt tctgtctcgg
     5401 cgcgtctgcg tctggctggc tggcataaat atctcactcg caatcaaatt cagccgatag
     5461 cggaacggga aggcgactgg agtgccatgt ccggttttca acaaaccatg caaatgctga
     5521 atgagggcat cgttcccact gcgatgctgg ttgccaacga tcagatggcg ctgggcgcaa
     5581 tgcgcgccat taccgagtcc gggctgcgcg ttggtgcgga tatctcggta gtgggatacg
     5641 acgataccga agacagctca tgttatatcc cgccgttaac caccatcaaa caggattttc
     5701 gcctgctggg gcaaaccagc gtggaccgct tgctgcaact ctctcagggc caggcggtga
     5761 agggcaatca gctgttgccc gtctcactgg tgaaaagaaa aaccaccctg gcgcccaata
     5821 cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt
     5881 cccgactgga aagcgggcag tgagcgcaac gcaattaatg taagttagct cactcattag
     5941 gcaccgggat ctcgaccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     6001 gcgcggggca tgactatcgt cgccgcactt atgactgtct tctttatcat gcaactcgta
     6061 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     6121 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     6181 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     6241 gccccacggg tgcgcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
     6301 tgccttact
//
