LOCUS       sequence-233746-        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: NK697
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    83..104
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(119..124,125..142,143..149)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    157..173
                     /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     162..184
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     181..197
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     181..197
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1813..1833
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             1920..1976
                     /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(4344..4361)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4344..4360)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4353..4375)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4569..4588)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4688..4710
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4748..4766)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4834..4938
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4939..5007,5008..5799)
                     /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(5157..5176)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5970..6558
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6459..6478
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6712..6729
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
       61 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      121 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      181 caggaaacag ctatgaccat gattacgcca agcttgtaac gactgcagtc ttgttgtcgt
      241 ttgtcgtcgt cgttctctgt ctcccaccca acttttgcca attgcaacag tgacttctgt
      301 acaatttgca acagatacca acgtgtcgag tcagcacaaa tggcgaggaa ggttcgcgtg
      361 cattgcttgc gcgcctcttt gtcatgggcg cacgctgatt tgtgtgtgat ttgcaccaaa
      421 tttcctgccg acacaacttg ggcccgtacc acacgacccg tttctggacc cgtttctggc
      481 ccagaaacgc gaatgttgcg aaagatgcga ccacgtgctc gctgtcgaca acacaaactc
      541 ccgcacaatc cccatgaaaa aaaaagtgct tgctgggctt gtgtctggcc tgtgtgcgtg
      601 gcgtggccgc gtgtggcttc gttcttgctt ctgctcactc agcctgttac acgcagagtg
      661 gctgcgagaa acaaggagag gggtttgcgg cattggcgca tgtgcatctg cctttttcat
      721 tgagtcagtc tgcagctcaa gaggttggtt tcgtcgacgg atatgagtga gcttctatgg
      781 atgatgagag gtggtggtgg ttgtatggtt ggtggttggt ggttggtgca aggcgagcca
      841 cgcgaagagc gacaaggatg agggagagga gaggagacta ggaaagaggt gaagggaagg
      901 gaagggcgag caatgacgac gagataaaag acagccacca ccgcacccgc acaaaagcat
      961 tgctcgcacg catggtcgtg tgttatgttt cacaccacca acataaccaa tacaagccat
     1021 caccaataca agccagcgct caagcacaca cgcacgcacg cacacacaat acacacgcac
     1081 gtgaaacgtt cacattgtcc gctcacaaat gcctttccct tctcgtcgct ttccttgcct
     1141 ttttttcttc ccctcctgac ttccttcctg ccttccttcc ttcctcccct tctccgtcca
     1201 ccaacaccat ttatttctct gctctttttc ctcgtgccac gtgagcagtc ttttcgcgtc
     1261 gtcttcttct ttgaccccgt ttaccgtcaa caccatcaac aaccagcaac atgtctggta
     1321 cggagtacaa gcctaccgtc cgcctcgcta cccgtgacga cgtgccccgt gctgtccgta
     1381 ccctcgccgc tgccttcgct gactaccctg ctacccgtca caccgtcgac cctgaccgcc
     1441 acatcgagcg tgtcaccgag ctgcaggagc tgttcctcac ccgcgtcggc ctcgacatcg
     1501 ggaaggtctg ggtcgccgac gacggtgctg ctgtcgctgt ctggaccacc cctgagtctg
     1561 tcgaggccgg tgctgtcttc gctgagatcg gcccccgcat ggctgagctg tctggctccc
     1621 gtctcgccgc tcagcagcag atggagggcc tcctcgcccc ccaccgcccc aaggagcccg
     1681 cctggttcct cgccaccgtc ggcgtctctc ctgaccacca gggcaagggt ctcggctctg
     1741 ccgtcgtcct ccctggcgtc gaggccgctg agcgtgctgg tgtccctgct ttcctcgaga
     1801 cctccgctcc tcgcaacctc cccttctacg agcgtctcgg cttcaccgtc accgccgacg
     1861 tggaggtccc cgagggtcct cgtacctggt gcatgacccg caagcccggt ggcagcggtg
     1921 ccaccaactt ttctctgctc aagcaggctg gcgacgttga agagaacccc ggacccatgg
     1981 tctccaaggg cgaggagacc acgatgggcg tcatcaagcc cgacatgaag atcaagctca
     2041 agatggaggg caacgtcaac ggccacgcct tcgtcatcga gggcgagggc gagggcaagc
     2101 cctacgacgg caccaacacc atcaacctcg aggtcaagga gggcgccccc ctccccttct
     2161 cctacgacat cctcaccacc gccttcgcct acggcaaccg cgccttcacc aagtaccccg
     2221 acgacatccc caactacttc aagcagtcct tccccgaggg ctactcctgg gagcgcacca
     2281 tgaccttcga ggacaagggc atcgtcaagg tcaagtccga catctctatg gaggaggact
     2341 ccttcatcta cgagatccac ctcaagggcg agaacttccc ccccaacggc cccgtcatgc
     2401 agaagaagac caccggctgg gacgcctcca ccgagcgcat gtacgtccgc gacggcgtcc
     2461 tcaagggcga cgtgaagcac aagctcctcc tcgagggcgg cggccaccac cgcgtcgact
     2521 tcaagaccat ctaccgcgcc aagaaggccg tcaagctccc cgactaccac ttcgtcgacc
     2581 accgcatcga gatcctcaac cacaacaagg actacaacaa ggtcaccgtc tacgagtccg
     2641 ccgtcgcccg caactccacc gacggcatgg acgagctgta caaggctggc ggtagcggtg
     2701 gctccatgtt gagggtacga ggaagatcgc tggaggcccg gtttgtgtac atcatcgtgc
     2761 tggctgtggt gtacacggcc acgtacatgt acagcgccac atccggtgac ccaagcgtac
     2821 cggttggcgg cgcagacatg ccgccgtggc tcgcacagct gggacgacca cgcgcagttg
     2881 cagaggagct ggcagaacag caacagctgc ttgacgcgac gcggcgcagc gacgacggag
     2941 ccaacagaca cagcaacgat ggcgccgccg cagtctcaca acaccagcag caccagcagc
     3001 aggcggagca cgatctggcc agcacacgga gtccaaagac accgacgcgg caagaggaca
     3061 aaggcgtgga cgagaaggag aaaggcatga agttcgaggt ggaggggggc gaagatggat
     3121 tcgtgcccag ggagcacatg aagaatgcga tgggtatgcc agacactgca gaggccgacc
     3181 cgtggaagcg gcgcaggcag gacaagtggc gcctcatcat cttgttccag cacaacttcc
     3241 ccatcctgaa gcaggcggta gaagggttcc agcgtgccag cgacatcatg gtgcccagca
     3301 tgatcatcgt cgacaactcc aaggacaggg atgcctcgaa cagcgtgtgg cttgtggagc
     3361 gggtggcaga ggtggtcgtg ccagaccgcc agctaaactt cccagagctg cacaactaca
     3421 tggccacgct ggcactcgag cgccggctcg aattctactt ctgggcccac gcagacaact
     3481 acgtgctgcc gctggagcct ggccgcgaca tgggcaagga cgccatcgac tgcctgcgcg
     3541 agcagatggc gcggttcccc aactggggca tgatcctctt cgcctacgac cacctggccg
     3601 cctaccgcac gcagacgcca gtccaggtgc catgggaccc acacgtgttc cagtacggct
     3661 cggagtgcga tgcgtatggg cgcatccgcg atgcgggcta cgacgcgcgc gcgtgcaaga
     3721 ttcactactc gtacgacatg aagcgggtga ttggcatcct ggacacggac tcgtacgacc
     3781 gggtcaaggc gaagctggac gaggaggcaa aggacaagtc caagcgcaac cagtggcgcg
     3841 agggagccat gtccgagaag gagcagcaat ggcgcaagcg catgaaggag gcgtcacgcg
     3901 catacttggc cgagaagtgg ggcgaggtga agtgcaagct gcgcggtgtg ccgtgccaca
     3961 agccctggcc ttactgccca acgtgcccca cagaccttcc ccagtgctac ggcaaggagg
     4021 cgacgtggga gcaactcgac cacattcacg ccagggtgca ccaggtgttt gacaatgatc
     4081 ccgacaagcc accaccgctc gaggcataag cgcctcgttg gtcattgcca ccttgttgcc
     4141 tcttttgctg ttgtcctcgt gtcgacgcgt gccacgtgcg cttgttgtga gttgattctg
     4201 cttcttgtgc acgtcatgcc gagcccttgg gccccgcaat aaacccacta tgccagtgta
     4261 actatccttc cccccaatgt ataactgtga cgatcaatga gcggtgcccg tctagaggat
     4321 ccccgggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4381 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4441 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4501 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     4561 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4621 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     4681 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     4741 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     4801 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4861 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4921 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     4981 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5041 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5101 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5161 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5221 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5281 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5341 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5401 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5461 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5521 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5581 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     5641 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     5701 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     5761 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     5821 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     5881 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     5941 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6001 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6061 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6121 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6181 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6241 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6301 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6361 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc
     6421 agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat
     6481 agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg
     6541 gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc
     6601 tggccttttg ctcacatctt ctttcctgcg ttatcccctg attctgtgga taaccgtatt
     6661 accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca
     6721 gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctc
//
