LOCUS       sequence-233698-        6097 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Reporter gene expression in S. rosetta.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6097)
  TITLE     King Lab plasmids
REFERENCE   2  (bases 1 to 6097)
  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: NK690
FEATURES             Location/Qualifiers
     source          1..6097
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     2..24
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(62..80)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        148..252
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(253..321,322..1113)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(471..490)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1284..1872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1773..1792
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2026..2043
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2160..2181
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2196..2201,2202..2219,2220..2226)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2234..2250
                     /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     2239..2261
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2258..2274
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2258..2274
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5755..5772)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5755..5771)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5764..5786)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5980..5999)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tccgggagct gcatgtgtca gaggttttca ccgtcatcac cgaaacgcgc gagacgaaag
       61 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
      121 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
      181 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
      241 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
      301 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
      361 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
      421 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
      481 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
      541 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
      601 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
      661 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
      721 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
      781 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
      841 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
      901 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
      961 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     1021 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     1081 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     1141 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     1201 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     1261 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     1321 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     1381 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     1441 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     1501 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     1561 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     1621 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     1681 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     1741 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     1801 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     1861 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     1921 tttgctcaca tcttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     1981 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     2041 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     2101 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     2161 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     2221 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     2281 tacgccaagc ttgtaacgac tgcagtcttg ttgtcgtttg tcgtcgtcgt tctctgtctc
     2341 ccacccaact tttgccaatt gcaacagtga cttctgtaca atttgcaaca gataccaacg
     2401 tgtcgagtca gcacaaatgg cgaggaaggt tcgcgtgcat tgcttgcgcg cctctttgtc
     2461 atgggcgcac gctgatttgt gtgtgatttg caccaaattt cctgccgaca caacttgggc
     2521 ccgtaccaca cgacccgttt ctggacccgt ttctggccca gaaacgcgaa tgttgcgaaa
     2581 gatgcgacca cgtgctcgct gtcgacaaca caaactcccg cacaatcccc atgaaaaaaa
     2641 aagtgcttgc tgggcttgtg tctggcctgt gtgcgtggcg tggccgcgtg tggcttcgtt
     2701 cttgcttctg ctcactcagc ctgttacacg cagagtggct gcgagaaaca aggagagggg
     2761 tttgcggcat tggcgcatgt gcatctgcct ttttcattga gtcagtctgc agctcaagag
     2821 gttggtttcg tcgacggata tgagtgagct tctatggatg atgagaggtg gtggtggttg
     2881 tatggttggt ggttggtggt tggtgcaagg cgagccacgc gaagagcgac aaggatgagg
     2941 gagaggagag gagactagga aagaggtgaa gggaagggaa gggcgagcaa tgacgacgag
     3001 ataaaagaca gccaccaccg cacccgcaca aaagcattgc tcgcacgcat ggtcgtgtgt
     3061 tatgtttcac accaccaaca taaccaatac aagccatcac caatacaagc cagcgctcaa
     3121 gcacacacgc acgcacgcac acacaataca cacgcacgtg aaacgttcac attgtccgct
     3181 cacaaatgcc tttcccttct cgtcgctttc cttgcctttt tttcttcccc tcctgacttc
     3241 cttcctgcct tccttccttc ctccccttct ccgtccacca acaccattta tttctctgct
     3301 ctttttcctc gtgccacgtg agcagtcttt tcgcgtcgtc ttcttctttg accccgttta
     3361 ccgtcaacac catcaacaac cagcaacatg ttgagggtac gaggaagatc gctggaggcc
     3421 cggtttgtgt acatcatcgt gctggctgtg gtgtacacgg ccacgtacat gtacagcgcc
     3481 acatccggtg acccaagcgt accggttggc ggcgcagaca tgccgccgtg gctcgcacag
     3541 ctgggacgac cacgcgcagt tgcagaggag ctggcagaac agcaacagct gcttgacgcg
     3601 acgcggcgca gcgacgacgg agccaacaga cacagcaacg atggcgccgc cgcagtctca
     3661 caacaccagc agcaccagca gcaggcggag cacgatctgg ccagcacacg gagtccaaag
     3721 acaccgacgc ggcaagagga caaaggcgtg gacgagaagg agaaaggcat gaagttcgag
     3781 gtggaggggg gcgaagatgg attcgtgccc agggagcaca tgaagaatgc gatgggtatg
     3841 ccagacactg cagaggccga cccgtggaag cggcgcaggc aggacaagtg gcgcctcatc
     3901 atcttgttcc agcacaactt ccccatcctg aagcaggcgg tagaagggtt ccagcgtgcc
     3961 agcgacatca tggtgcccag catgatcatc gtcgacaact ccaaggacag ggatgcctcg
     4021 aacagcgtgt ggcttgtgga gcgggtggca gaggtggtcg tgccagaccg ccagctaaac
     4081 ttcccagagc tgcacaacta catggccacg ctggcactcg agcgccggct cgaattctac
     4141 ttctgggccc acgcagacaa ctacgtgctg ccgctggagc ctggccgcga catgggcaag
     4201 gacgccatcg actgcctgcg cgagcagatg gcgcggttcc ccaactgggg catgatcctc
     4261 ttcgcctacg accacctggc cgcctaccgc acgcagacgc tagtccaggt gccatgggac
     4321 ccacacgtgt tccagtacgg ctcggagtgc gatgcgtatg ggcgcatccg cgatgcgggc
     4381 tacgacgcgc gcgcgtgcaa gattcactac tcgtacgaca tgaagcgggt gattggcatc
     4441 ctggacacgg actcgtacga ccgggtcaag gcgaagctgg acgaggaggc aaaggacaag
     4501 tccaagcgca accagtggcg cgagggagcc atgtccgaga aggagcagca atggcgcaag
     4561 cgcatgaagg aggcgtcacg cgcatacttg gccgagaagt ggggcgaggt gaagtgcaag
     4621 ctgcgcggtg tgccgtgcca caagccctgg ccttactgcc caacgtgccc cacagacctt
     4681 ccccagtgct acggcaagga ggcgacgtgg gagcaactcg accacattca cgccagggtg
     4741 caccaggtgt ttgacaatga tcccgacaag ccaccaccgc tcgaggcagg cggtagcggt
     4801 ggctccatgg tctccaaggg cgaggagacc acgatgggcg tcatcaagcc cgacatgaag
     4861 atcaagctca agatggaggg caacgtcaac ggccacgcct tcgtcatcga gggcgagggc
     4921 gagggcaagc cctacgacgg caccaacacc atcaaccttg aggtcaaaga aggtgctcct
     4981 ctgcccttta gctacgacat cctcaccacc gccttctcct acggcaaccg cgccttcacc
     5041 aagtaccccg acgacatccc caactacttc aagcagtcct tccccgaggg ctactcctgg
     5101 gagcgcacca tgaccttcga ggacaagggc atcgtcaagg tcaagtccga catctctatg
     5161 gaggaggact ccttcatcta cgagatccac ctcaagggcg agaacttccc tcccaacggc
     5221 cccgtcatgc agaaggagac tacgggttgg gacgcctcca ccgagcgcat gtacgtccgc
     5281 gacggtgttc tcaagggcga cgtgaagatg aaacttctcc ttgagggcgg cggccaccac
     5341 cgcgtcgact tcaagaccat ctaccgcgcc aagaaggccg tcaagctccc cgactaccac
     5401 ttcgtcgacc accgcatcga gatcctcaac cacgacaagg actacaacaa agtcactgtc
     5461 tacgagatcg ccgtcgcccg caactctacc gacggcatgg acgagcttta caaggcttaa
     5521 gcgcctcgtt ggtcattgcc accttgttgc ctcttttgct gttgtcctcg tgtcgacgcg
     5581 tgccacgtgc gcttgttgtg agttgattct gcttcttgtg cacgtcatgc cgagcccttg
     5641 ggccccgcaa taaacccact atgccagtgt aactatcctt ccccccaatg tataactgtg
     5701 acgatcaatg agcggtgccc gtctagagga tccccgggta ccgagctcga attcactggc
     5761 cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt acccaactta atcgccttgc
     5821 agcacatccc cctttcgcca gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc
     5881 ccaacagttg cgcagcctga atggcgaatg gcgcctgatg cggtattttc tccttacgca
     5941 tctgtgcggt atttcacacc gcatatggtg cactctcagt acaatctgct ctgatgccgc
     6001 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct
     6061 gctcccggca tccgcttaca gacaagctgt gaccgtc
//
