LOCUS       sequence-160476-        7137 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  expresses E. coli succinyl-CoA synthetase.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7137)
  TITLE     pSucCD2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7137)
  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"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pSucCD2
FEATURES             Location/Qualifiers
     source          1..7137
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(26..73)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     69..87
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     CDS             complement(150..173)
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=8xHis
                     /translation="HHHHHHHH"
     RBS             2307..2329
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    2344..2368
                     /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(2368..2387)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2369..2387)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2475..2494)
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     promoter        2700..2777
                     /gene="lacI"
                     /label=lacI promoter
     CDS             2778..3860
                     /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(2797..2816)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     CDS             4432..4623
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(4617..4639)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    4725..4867
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     4780..4799
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4882..4899)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5053..5641)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5133..5152)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(5752..5771)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             5763..6578
                     /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             5763..6578
                     /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(5836..5855)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      complement(6671..7126)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6808..6829)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7020..7039
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 atccggatat agttcctcct ttcagcaaaa aacccctcaa gacccgttta gaggccccaa
       61 ggggttatgc tagttattgc tcagcggtgg cagcagccaa ctcagcttcc tttcgggctt
      121 tgtttagcag cctaggtatt aatcaattag tggtggtggt ggtggtggtg gtgctcgagt
      181 gcggccgcaa gcttgtcgac ggagctcgcc tgcaggcgcg ccaaggcctg tacagaattc
      241 actagtgatt gagcatcgag ctcttatttc agaacagttt tcagtgcttc accgatatcc
      301 gccaggctgc gaacggtttt cacgcctgcg gcttccagag cagcgaattt ctcatccgca
      361 gtccctttcc caccggcaat gatggcaccc gcgtggccca tacgtttgcc tttcggcgca
      421 gttacaccag cgatgtaacc cacaactggc ttggtaacgt gctctttgat gtacgcagct
      481 gcttcttctt cagcgctacc gccgatctca ccgatcatca cgatcgcttc ggtctgcgga
      541 tctttttcga acatttcgag gatgtcgata aagttagagc cagggatcgg gtcaccgccg
      601 ataccgacac aggtcgactg accgaaaccg taatccgtgg tctgtttaac cgcttcatag
      661 gtcagtgtac cggaacggga aacgataccc actttacccg gtttgtgaat gtgaccaggc
      721 tgaataccga ttttgcattc acccggagtg ataacgcctg ggcagttcgg gccgatcata
      781 cgaacgcctg cttcatccag cttcactttc acggtcagca tatccagcgt cgggatgcct
      841 tcagtgatgg tgataatcag tttgatgcct gcgtcgatgg cttccagaat ggagtctttg
      901 cagaacggtg ctggtacgta gataacagaa gcggtagcgc cagtagcagc aacagcttca
      961 cgcacggtgt tgaacaccgg caggccgagg tgggtagtac cgcctttacc tggggttacg
     1021 ccgccaacca ttttagtgcc gtatgcaatg gcctgttctg agtggaaagt cccctggcta
     1081 ccggtaaagc cctggcagat aaccttggtg tttttatcaa ttaaaatgga cattatttcc
     1141 cctccactgc ggcaacaacc tgctgagctg catccgtcag accttttgct gcaataatat
     1201 tcaggccgct gtcagccagt ttcttcgcgc cgagttcggc gttgttacct tccagacgta
     1261 ccacgaccgg tacgttaaca cccacttccg ctaccgcacc gatgataccg tcagcgatca
     1321 ggtcgcaacg aacgataccg ccgaagatgt taaccagaac ggctttcact ttgtcgtcag
     1381 agaggatgat tttgaacgct tcggttacac gttctttggt tgcgccgccg ccaacgtcca
     1441 ggaagttagc cggttcgccg ccgtgcagtt taacgatgtc catcgtaccc atcgccaggc
     1501 ctgcgccgtt aaccatacaa ccgatgttac cgtccagcgc aacgtagttc agttcccact
     1561 gtgcagcctg tgcttcacgc ggatcttcct gcgactggtc acgcatttcg cgcagatcag
     1621 gctggcggaa cagggcgttg ccatcagcgc ccagtttgcc gtcgaggcaa atcagatcgc
     1681 cctgtttggt gatgaccagc gggttgattt cgatcagcgc caggtcgcgc tccaggaaaa
     1741 tggtcgccag gcccatgaag attttggtga actgctgaac cagtttacct tccagaccca
     1801 gtttgaacgc cagctcgcgt ccctgatacg gcatcgggcc agtcagcgga tcaagcgcaa
     1861 ctttgtggat caggtgcgga gtttcttccg ccactttttc gatttccacg ccgccttcgg
     1921 tggaggccat aaagaccacg cgacgtgaac tacggtctac aacggcaccg agatacagct
     1981 ctttagcgat atcggtcgct gcttcaacca gaatctggtt aaccggttgg ccattggcat
     2041 ctgtttgata cgttaccaga cgcttgccca gccagttttc tgcaaaagca cggatgtctt
     2101 ctttgctgtt tacaactttc acaccgcccg ctttaccgcg gccgcctgcg tgaacctgac
     2161 atttcactac ccacggaccg gcaccgattt ttgaagcggc ttcttctgct tcgcgcggag
     2221 tagtacaggc ataacccacc ggtgctggta agccatagcg ggcaaacagt tgttttgcct
     2281 gatattcatg taagttcata tgtatatctc cttcttaaag ttaaacaaaa ttatttctag
     2341 aggggaattg ttatccgctc acaattcccc tatagtgagt cgtattaatt tcgcgggatc
     2401 gagatcgatc tcgatcctct acgccggacg catcgtggcc ggcatcaccg gcgccacagg
     2461 tgcggttgct ggcgcctata tcgccgacat caccgatggg gaagatcggg ctcgccactt
     2521 cgggctcatg agcgcttgtt tcggcgtggg tatggtggca ggccccgtgg ccgggggact
     2581 gttgggcgcc atctccttgc atgcaccatt ccttgcggcg gcggtgctca acggcctcaa
     2641 cctactactg ggctgcttcc taatgcagga gtcgcataag ggagagcgtc gagatcccgg
     2701 acaccatcga atggcgcaaa acctttcgcg gtatggcatg atagcgcccg gaagagagtc
     2761 aattcagggt ggtgaatgtg aaaccagtaa cgttatacga tgtcgcagag tatgccggtg
     2821 tctcttatca gaccgtttcc cgcgtggtga accaggccag ccacgtttct gcgaaaacgc
     2881 gggaaaaagt ggaagcggcg atggcggagc tgaattacat tcccaaccgc gtggcacaac
     2941 aactggcggg caaacagtcg ttgctgattg gcgttgccac ctccagtctg gccctgcacg
     3001 cgccgtcgca aattgtcgcg gcgattaaat ctcgcgccga tcaactgggt gccagcgtgg
     3061 tggtgtcgat ggtagaacga agcggcgtcg aagcctgtaa agcggcggtg cacaatcttc
     3121 tcgcgcaacg cgtcagtggg ctgatcatta actatccgct ggatgaccag gatgccattg
     3181 ctgtggaagc tgcctgcact aatgttccgg cgttatttct tgatgtctct gaccagacac
     3241 ccatcaacag tattattttc tcccatgaag acggtacgcg actgggcgtg gagcatctgg
     3301 tcgcattggg tcaccagcaa atcgcgctgt tagcgggccc attaagttct gtctcggcgc
     3361 gtctgcgtct ggctggctgg cataaatatc tcactcgcaa tcaaattcag ccgatagcgg
     3421 aacgggaagg cgactggagt gccatgtccg gttttcaaca aaccatgcaa atgctgaatg
     3481 agggcatcgt tcccactgcg atgctggttg ccaacgatca gatggcgctg ggcgcaatgc
     3541 gcgccattac cgagtccggg ctgcgcgttg gtgcggacat ctcggtagtg ggatacgacg
     3601 ataccgaaga cagctcatgt tatatcccgc cgttaaccac catcaaacag gattttcgcc
     3661 tgctggggca aaccagcgtg gaccgcttgc tgcaactctc tcagggccag gcggtgaagg
     3721 gcaatcagct gttgcccgtc tcactggtga aaagaaaaac caccctggcg cccaatacgc
     3781 aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc
     3841 gactggaaag cgggcagtga gcgcaacgca attaatgtaa gttagctcac tcattaggca
     3901 ccgggatctc gaccgatgcc cttgagagcc ttcaacccag tcagctcctt ccggtgggcg
     3961 cggggcatga ctagcatgat cgtgctcctg tcgttgagga cccggctagg ctggcggggt
     4021 tgccttactg gttagcagaa tgaatcaccg atacgcgagc gaacgtgaag cgactgctgc
     4081 tgcaaaacgt ctgcgacctg agcaacaaca tgaatggtct tcggtttccg tgtttcgtaa
     4141 agtctggaaa cgcggaagtc agcgccctgc accattatgt tccggatctg catcgcagga
     4201 tgctgctggc taccctgtgg aacacctaca tctgtattaa cgaagcgctg gcattgaccc
     4261 tgagtgattt ttctctggtc ccgccgcatc cataccgcca gttgtttacc ctcacaacgt
     4321 tccagtaacc gggcatgttc atcatcagta acccgtatcg tgagcatcct ctctcgtttc
     4381 atcggtatca ttacccccat gaacagaaat cccccttaca cggaggcatc agtgaccaaa
     4441 caggaaaaaa ccgcccttaa catggcccgc tttatcagaa gccagacatt aacgcttctg
     4501 gagaaactca acgagctgga cgcggatgaa caggcagaca tctgtgaatc gcttcacgac
     4561 cacgctgatg agctttaccg cagctgcctc gcgcgtttcg gtgatgacgg tgaaaacctc
     4621 tgacacatgc agctcccgga gacggtcaca gcttgtctgt aagcggatgc cgggagcaga
     4681 caagcccgtc agggcgcgtc agcgggtgtt ggcgggtgtc ggggcgcagc catgacccag
     4741 tcacgtagcg atagcggagt gtatactggc ttaactatgc ggcatcagag cagattgtac
     4801 tgagagtgca ccatatatgc ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg
     4861 catcaggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg
     4921 gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa
     4981 cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc
     5041 gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc
     5101 aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag
     5161 ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct
     5221 cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta
     5281 ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc
     5341 cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc
     5401 agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt
     5461 gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct
     5521 gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc
     5581 tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca
     5641 agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta
     5701 agggattttg gtcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg
     5761 ttatgagcca tattcaacgg gaaacgtctt gctctaggcc gcgattaaat tccaacatgg
     5821 atgctgattt atatgggtat aaatgggctc gcgataatgt cgggcaatca ggtgcgacaa
     5881 tctatcgatt gtatgggaag cccgatgcgc cagagttgtt tctgaaacat ggcaaaggta
     5941 gcgttgccaa tgatgttaca gatgagatgg tcagactaaa ctggctgacg gaatttatgc
     6001 ctcttccgac catcaagcat tttatccgta ctcctgatga tgcatggtta ctcaccactg
     6061 cgatccccgg gaaaacagca ttccaggtat tagaagaata tcctgattca ggtgaaaata
     6121 ttgttgatgc gctggcagtg ttcctgcgcc ggttgcattc gattcctgtt tgtaattgtc
     6181 cttttaacag cgatcgcgta tttcgtctcg ctcaggcgca atcacgaatg aataacggtt
     6241 tggttgatgc gagtgatttt gatgacgagc gtaatggctg gcctgttgaa caagtctgga
     6301 aagaaatgca taaacttttg ccattctcac cggattcagt cgtcactcat ggtgatttct
     6361 cacttgataa ccttattttt gacgagggga aattaatagg ttgtattgat gttggacgag
     6421 tcggaatcgc agaccgatac caggatcttg ccatcctatg gaactgcctc ggtgagtttt
     6481 ctccttcatt acagaaacgg ctttttcaaa aatatggtat tgataatcct gatatgaata
     6541 aattgcagtt tcatttgatg ctcgatgagt ttttctaaga attaattcat gagcggatac
     6601 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6661 gtgccacctg aaattgtaaa cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
     6721 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
     6781 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
     6841 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
     6901 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
     6961 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
     7021 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
     7081 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgcca
//
