LOCUS       sequence-233742-        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: NK694
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     63..82
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     316..333
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    450..471
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(486..491,492..509,510..516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    524..540
                     /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     529..551
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     548..564
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     548..564
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2180..2200
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2287..2343
                     /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(4711..4728)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4711..4727)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4720..4742)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4936..4955)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5055..5077
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5115..5133)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5201..5305
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5306..5374,5375..6166)
                     /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(5524..5543)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6337..6763,1..162)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
       61 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
      121 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      181 tttttacggt tcctggcctt ttgctggcct tttgctcaca tcttctttcc tgcgttatcc
      241 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
      301 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
      361 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
      421 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
      481 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
      541 tttcacacag gaaacagcta tgaccatgat tacgccaagc ttgtaacgac tgcagtcttg
      601 ttgtcgtttg tcgtcgtcgt tctctgtctc ccacccaact tttgccaatt gcaacagtga
      661 cttctgtaca atttgcaaca gataccaacg tgtcgagtca gcacaaatgg cgaggaaggt
      721 tcgcgtgcat tgcttgcgcg cctctttgtc atgggcgcac gctgatttgt gtgtgatttg
      781 caccaaattt cctgccgaca caacttgggc ccgtaccaca cgacccgttt ctggacccgt
      841 ttctggccca gaaacgcgaa tgttgcgaaa gatgcgacca cgtgctcgct gtcgacaaca
      901 caaactcccg cacaatcccc atgaaaaaaa aagtgcttgc tgggcttgtg tctggcctgt
      961 gtgcgtggcg tggccgcgtg tggcttcgtt cttgcttctg ctcactcagc ctgttacacg
     1021 cagagtggct gcgagaaaca aggagagggg tttgcggcat tggcgcatgt gcatctgcct
     1081 ttttcattga gtcagtctgc agctcaagag gttggtttcg tcgacggata tgagtgagct
     1141 tctatggatg atgagaggtg gtggtggttg tatggttggt ggttggtggt tggtgcaagg
     1201 cgagccacgc gaagagcgac aaggatgagg gagaggagag gagactagga aagaggtgaa
     1261 gggaagggaa gggcgagcaa tgacgacgag ataaaagaca gccaccaccg cacccgcaca
     1321 aaagcattgc tcgcacgcat ggtcgtgtgt tatgtttcac accaccaaca taaccaatac
     1381 aagccatcac caatacaagc cagcgctcaa gcacacacgc acgcacgcac acacaataca
     1441 cacgcacgtg aaacgttcac attgtccgct cacaaatgcc tttcccttct cgtcgctttc
     1501 cttgcctttt tttcttcccc tcctgacttc cttcctgcct tccttccttc ctccccttct
     1561 ccgtccacca acaccattta tttctctgct ctttttcctc gtgccacgtg agcagtcttt
     1621 tcgcgtcgtc ttcttctttg accccgttta ccgtcaacac catcaacaac cagcaacatg
     1681 tctggtacgg agtacaagcc taccgtccgc ctcgctaccc gtgacgacgt gccccgtgct
     1741 gtccgtaccc tcgccgctgc cttcgctgac taccctgcta cccgtcacac cgtcgaccct
     1801 gaccgccaca tcgagcgtgt caccgagctg caggagctgt tcctcacccg cgtcggcctc
     1861 gacatcggga aggtctgggt cgccgacgac ggtgctgctg tcgctgtctg gaccacccct
     1921 gagtctgtcg aggccggtgc tgtcttcgct gagatcggcc cccgcatggc tgagctgtct
     1981 ggctcccgtc tcgccgctca gcagcagatg gagggcctcc tcgcccccca ccgccccaag
     2041 gagcccgcct ggttcctcgc caccgtcggc gtctctcctg accaccaggg caagggtctc
     2101 ggctctgccg tcgtcctccc tggcgtcgag gccgctgagc gtgctggtgt ccctgctttc
     2161 ctcgagacct ccgctcctcg caacctcccc ttctacgagc gtctcggctt caccgtcacc
     2221 gccgacgtgg aggtccccga gggtcctcgt acctggtgca tgacccgcaa gcccggtggc
     2281 agcggtgcca ccaacttttc tctgctcaag caggctggcg acgttgaaga gaaccccgga
     2341 cccatgttga gggtacgagg aagatcgctg gaggcccggt ttgtgtacat catcgtgctg
     2401 gctgtggtgt acacggccac gtacatgtac agcgccacat ccggtgaccc aagcgtaccg
     2461 gttggcggcg cagacatgcc gccgtggctc gcacagctgg gacgaccacg cgcagttgca
     2521 gaggagctgg cagaacagca acagctgctt gacgcgacgc ggcgcagcga cgacggagcc
     2581 aacagacaca gcaacgatgg cgccgccgca gtctcacaac accagcagca ccagcagcag
     2641 gcggagcacg atctggccag cacacggagt ccaaagacac cgacgcggca agaggacaaa
     2701 ggcgtggacg agaaggagaa aggcatgaag ttcgaggtgg aggggggcga agatggattc
     2761 gtgcccaggg agcacatgaa gaatgcgatg ggtatgccag acactgcaga ggccgacccg
     2821 tggaagcggc gcaggcagga caagtggcgc ctcatcatct tgttccagca caacttcccc
     2881 atcctgaagc aggcggtaga agggttccag cgtgccagcg acatcatggt gcccagcatg
     2941 atcatcgtcg acaactccaa ggacagggat gcctcgaaca gcgtgtggct tgtggagcgg
     3001 gtggcagagg tggtcgtgcc agaccgccag ctaaacttcc cagagctgca caactacatg
     3061 gccacgctgg cactcgagcg ccggctcgaa ttctacttct gggcccacgc agacaactac
     3121 gtgctgccgc tggagcctgg ccgcgacatg ggcaaggacg ccatcgactg cctgcgcgag
     3181 cagatggcgc ggttccccaa ctggggcatg atcctcttcg cctacgacca cctggccgcc
     3241 taccgcacgc agacgctagt ccaggtgcca tgggacccac acgtgttcca gtacggctcg
     3301 gagtgcgatg cgtatgggcg catccgcgat gcgggctacg acgcgcgcgc gtgcaagatt
     3361 cactactcgt acgacatgaa gcgggtgatt ggcatcctgg acacggactc gtacgaccgg
     3421 gtcaaggcga agctggacga ggaggcaaag gacaagtcca agcgcaacca gtggcgcgag
     3481 ggagccatgt ccgagaagga gcagcaatgg cgcaagcgca tgaaggaggc gtcacgcgca
     3541 tacttggccg agaagtgggg cgaggtgaag tgcaagctgc gcggtgtgcc gtgccacaag
     3601 ccctggcctt actgcccaac gtgccccaca gaccttcccc agtgctacgg caaggaggcg
     3661 acgtgggagc aactcgacca cattcacgcc agggtgcacc aggtgtttga caatgatccc
     3721 gacaagccac caccgctcga ggcaggcggt agcggtggct ccatggtctc caagggcgag
     3781 gagaccacga tgggcgtcat caagcccgac atgaagatca agctcaagat ggagggcaac
     3841 gtcaacggcc acgccttcgt catcgagggc gagggcgagg gcaagcccta cgacggcacc
     3901 aacaccatca acctcgaggt caaggagggc gcccccctcc ccttctccta cgacatcctc
     3961 accaccgcct tcgcctacgg caaccgcgcc ttcaccaagt accccgacga catccccaac
     4021 tacttcaagc agtccttccc cgagggctac tcctgggagc gcaccatgac cttcgaggac
     4081 aagggcatcg tcaaggtcaa gtccgacatc tctatggagg aggactcctt catctacgag
     4141 atccacctca agggcgagaa cttccccccc aacggccccg tcatgcagaa gaagaccacc
     4201 ggctgggacg cctccaccga gcgcatgtac gtccgcgacg gcgtcctcaa gggcgacgtg
     4261 aagcacaagc tcctcctcga gggcggcggc caccaccgcg tcgacttcaa gaccatctac
     4321 cgcgccaaga aggccgtcaa gctccccgac taccacttcg tcgaccaccg catcgagatc
     4381 ctcaaccaca acaaggacta caacaaggtc accgtctacg agtccgccgt cgcccgcaac
     4441 tccaccgacg gcatggacga gctgtacaag gcttaagcgc ctcgttggtc attgccacct
     4501 tgttgcctct tttgctgttg tcctcgtgtc gacgcgtgcc acgtgcgctt gttgtgagtt
     4561 gattctgctt cttgtgcacg tcatgccgag cccttgggcc ccgcaataaa cccactatgc
     4621 cagtgtaact atccttcccc ccaatgtata actgtgacga tcaatgagcg gtgcccgtct
     4681 agaggatccc cgggtaccga gctcgaattc actggccgtc gttttacaac gtcgtgactg
     4741 ggaaaaccct ggcgttaccc aacttaatcg ccttgcagca catccccctt tcgccagctg
     4801 gcgtaatagc gaagaggccc gcaccgatcg cccttcccaa cagttgcgca gcctgaatgg
     4861 cgaatggcgc ctgatgcggt attttctcct tacgcatctg tgcggtattt cacaccgcat
     4921 atggtgcact ctcagtacaa tctgctctga tgccgcatag ttaagccagc cccgacaccc
     4981 gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc ccggcatccg cttacagaca
     5041 agctgtgacc gtctccggga gctgcatgtg tcagaggttt tcaccgtcat caccgaaacg
     5101 cgcgagacga aagggcctcg tgatacgcct atttttatag gttaatgtca tgataataat
     5161 ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt
     5221 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     5281 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     5341 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     5401 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5461 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5521 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5581 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5641 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5701 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5761 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5821 aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc gcaaactatt
     5881 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5941 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     6001 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     6061 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     6121 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     6181 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     6241 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     6301 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     6361 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     6421 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     6481 tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     6541 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     6601 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     6661 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     6721 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aag
//
