LOCUS       sequence-233744-        6763 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 6763)
  TITLE     King Lab plasmids
REFERENCE   2  (bases 1 to 6763)
  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: NK695
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    89..110
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(125..130,131..148,149..155)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    163..179
                     /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     168..190
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     187..203
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     187..203
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1819..1839
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1926..1982
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     primer_bind     complement(4350..4367)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4350..4366)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4359..4381)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4575..4594)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4694..4716
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4754..4772)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4840..4944
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4945..5013,5014..5805)
                     /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(5163..5182)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5976..6564
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6465..6484
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6718..6735
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
       61 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
      121 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
      181 ttcacacagg aaacagctat gaccatgatt acgccaagct tgtaacgact gcagtcttgt
      241 tgtcgtttgt cgtcgtcgtt ctctgtctcc cacccaactt ttgccaattg caacagtgac
      301 ttctgtacaa tttgcaacag ataccaacgt gtcgagtcag cacaaatggc gaggaaggtt
      361 cgcgtgcatt gcttgcgcgc ctctttgtca tgggcgcacg ctgatttgtg tgtgatttgc
      421 accaaatttc ctgccgacac aacttgggcc cgtaccacac gacccgtttc tggacccgtt
      481 tctggcccag aaacgcgaat gttgcgaaag atgcgaccac gtgctcgctg tcgacaacac
      541 aaactcccgc acaatcccca tgaaaaaaaa agtgcttgct gggcttgtgt ctggcctgtg
      601 tgcgtggcgt ggccgcgtgt ggcttcgttc ttgcttctgc tcactcagcc tgttacacgc
      661 agagtggctg cgagaaacaa ggagaggggt ttgcggcatt ggcgcatgtg catctgcctt
      721 tttcattgag tcagtctgca gctcaagagg ttggtttcgt cgacggatat gagtgagctt
      781 ctatggatga tgagaggtgg tggtggttgt atggttggtg gttggtggtt ggtgcaaggc
      841 gagccacgcg aagagcgaca aggatgaggg agaggagagg agactaggaa agaggtgaag
      901 ggaagggaag ggcgagcaat gacgacgaga taaaagacag ccaccaccgc acccgcacaa
      961 aagcattgct cgcacgcatg gtcgtgtgtt atgtttcaca ccaccaacat aaccaataca
     1021 agccatcacc aatacaagcc agcgctcaag cacacacgca cgcacgcaca cacaatacac
     1081 acgcacgtga aacgttcaca ttgtccgctc acaaatgcct ttcccttctc gtcgctttcc
     1141 ttgccttttt ttcttcccct cctgacttcc ttcctgcctt ccttccttcc tccccttctc
     1201 cgtccaccaa caccatttat ttctctgctc tttttcctcg tgccacgtga gcagtctttt
     1261 cgcgtcgtct tcttctttga ccccgtttac cgtcaacacc atcaacaacc agcaacatgt
     1321 ctggtacgga gtacaagcct accgtccgcc tcgctacccg tgacgacgtg ccccgtgctg
     1381 tccgtaccct cgccgctgcc ttcgctgact accctgctac ccgtcacacc gtcgaccctg
     1441 accgccacat cgagcgtgtc accgagctgc aggagctgtt cctcacccgc gtcggcctcg
     1501 acatcgggaa ggtctgggtc gccgacgacg gtgctgctgt cgctgtctgg accacccctg
     1561 agtctgtcga ggccggtgct gtcttcgctg agatcggccc ccgcatggct gagctgtctg
     1621 gctcccgtct cgccgctcag cagcagatgg agggcctcct cgccccccac cgccccaagg
     1681 agcccgcctg gttcctcgcc accgtcggcg tctctcctga ccaccagggc aagggtctcg
     1741 gctctgccgt cgtcctccct ggcgtcgagg ccgctgagcg tgctggtgtc cctgctttcc
     1801 tcgagacctc cgctcctcgc aacctcccct tctacgagcg tctcggcttc accgtcaccg
     1861 ccgacgtgga ggtccccgag ggtcctcgta cctggtgcat gacccgcaag cccggtggca
     1921 gcggtgccac caacttttct ctgctcaagc aggctggcga cgttgaagag aaccccggac
     1981 ccatgttgag ggtacgagga agatcgctgg aggcccggtt tgtgtacatc atcgtgctgg
     2041 ctgtggtgta cacggccacg tacatgtaca gcgccacatc cggtgaccca agcgtaccgg
     2101 ttggcggcgc agacatgccg ccgtggctcg cacagctggg acgaccacgc gcagttgcag
     2161 aggagctggc agaacagcaa cagctgcttg acgcgacgcg gcgcagcgac gacggagcca
     2221 acagacacag caacgatggc gccgccgcag tctcacaaca ccagcagcac cagcagcagg
     2281 cggagcacga tctggccagc acacggagtc caaagacacc gacgcggcaa gaggacaaag
     2341 gcgtggacga gaaggagaaa ggcatgaagt tcgaggtgga ggggggcgaa gatggattcg
     2401 tgcccaggga gcacatgaag aatgcgatgg gtatgccaga cactgcagag gccgacccgt
     2461 ggaagcggcg caggcaggac aagtggcgcc tcatcatctt gttccagcac aacttcccca
     2521 tcctgaagca ggcggtagaa gggttccagc gtgccagcga catcatggtg cccagcatga
     2581 tcatcgtcga caactccaag gacagggatg cctcgaacag cgtgtggctt gtggagcggg
     2641 tggcagaggt ggtcgtgcca gaccgccagc taaacttccc agagctgcac aactacatgg
     2701 ccacgctggc actcgagcgc cggctcgaat tctacttctg ggcccacgca gacaactacg
     2761 tgctgccgct ggagcctggc cgcgacatgg gcaaggacgc catcgactgc ctgcgcgagc
     2821 agatggcgcg gttccccaac tggggcatga tcctcttcgc ctacgaccac ctggccgcct
     2881 accgcacgca gacgccagtc caggtgccat gggacccaca cgtgttccag tacggctcgg
     2941 agtgcgatgc gtatgggcgc atccgcgatg cgggctacga cgcgcgcgcg tgcaagattc
     3001 actactcgta cgacatgaag cgggtgattg gcatcctgga cacggactcg tacgaccggg
     3061 tcaaggcgaa gctggacgag gaggcaaagg acaagtccaa gcgcaaccag tggcgcgagg
     3121 gagccatgtc cgagaaggag cagcaatggc gcaagcgcat gaaggaggcg tcacgcgcat
     3181 acttggccga gaagtggggc gaggtgaagt gcaagctgcg cggtgtgccg tgccacaagc
     3241 cctggcctta ctgcccaacg tgccccacag accttcccca gtgctacggc aaggaggcga
     3301 cgtgggagca actcgaccac attcacgcca gggtgcacca ggtgtttgac aatgatcccg
     3361 acaagccacc accgctcgag gcaggcggta gcggtggctc catggtctcc aagggcgagg
     3421 agaccacgat gggcgtcatc aagcccgaca tgaagatcaa gctcaagatg gagggcaacg
     3481 tcaacggcca cgccttcgtc atcgagggcg agggcgaggg caagccctac gacggcacca
     3541 acaccatcaa cctcgaggtc aaggagggcg cccccctccc cttctcctac gacatcctca
     3601 ccaccgcctt cgcctacggc aaccgcgcct tcaccaagta ccccgacgac atccccaact
     3661 acttcaagca gtccttcccc gagggctact cctgggagcg caccatgacc ttcgaggaca
     3721 agggcatcgt caaggtcaag tccgacatct ctatggagga ggactccttc atctacgaga
     3781 tccacctcaa gggcgagaac ttccccccca acggccccgt catgcagaag aagaccaccg
     3841 gctgggacgc ctccaccgag cgcatgtacg tccgcgacgg cgtcctcaag ggcgacgtga
     3901 agcacaagct cctcctcgag ggcggcggcc accaccgcgt cgacttcaag accatctacc
     3961 gcgccaagaa ggccgtcaag ctccccgact accacttcgt cgaccaccgc atcgagatcc
     4021 tcaaccacaa caaggactac aacaaggtca ccgtctacga gtccgccgtc gcccgcaact
     4081 ccaccgacgg catggacgag ctgtacaagg cttaagcgcc tcgttggtca ttgccacctt
     4141 gttgcctctt ttgctgttgt cctcgtgtcg acgcgtgcca cgtgcgcttg ttgtgagttg
     4201 attctgcttc ttgtgcacgt catgccgagc ccttgggccc cgcaataaac ccactatgcc
     4261 agtgtaacta tccttccccc caatgtataa ctgtgacgat caatgagcgg tgcccgtcta
     4321 gaggatcccc gggtaccgag ctcgaattca ctggccgtcg ttttacaacg tcgtgactgg
     4381 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     4441 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgcag cctgaatggc
     4501 gaatggcgcc tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata
     4561 tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagcc ccgacacccg
     4621 ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc cggcatccgc ttacagacaa
     4681 gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt caccgtcatc accgaaacgc
     4741 gcgagacgaa agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg
     4801 gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta
     4861 tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt
     4921 caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc
     4981 ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa
     5041 gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt
     5101 aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt
     5161 ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc
     5221 atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg
     5281 gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg
     5341 gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac
     5401 atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca
     5461 aacgacgagc gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta
     5521 actggcgaac tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat
     5581 aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa
     5641 tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag
     5701 ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat
     5761 agacagatcg ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt
     5821 tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg
     5881 aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga
     5941 gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta
     6001 atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa
     6061 gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact
     6121 gttcttctag tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca
     6181 tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt
     6241 accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg
     6301 ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag
     6361 cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta
     6421 agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat
     6481 ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg
     6541 tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc
     6601 ttttgctggc cttttgctca catcttcttt cctgcgttat cccctgattc tgtggataac
     6661 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
     6721 gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca aac
//
