LOCUS       sequence-229610-        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"
     primer_bind     complement(10..28)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        96..200
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(201..269,270..1061)
                     /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(419..438)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1232..1820
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1721..1740
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     1974..1991
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2108..2129
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2144..2149,2150..2167,2168..2174)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2182..2198
                     /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     2187..2209
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2206..2222
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2206..2222
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3838..3858
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             3945..4001
                     /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(6369..6386)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6369..6385)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6378..6400)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6594..6613)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6713..6735
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
ORIGIN
        1 gacgaaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt
       61 cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt
      121 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
      181 aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt
      241 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
      301 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
      361 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
      421 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
      481 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
      541 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
      601 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
      661 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
      721 acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg
      781 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
      841 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg
      901 gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct
      961 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     1021 agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac caagtttact
     1081 catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga
     1141 tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt
     1201 cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct
     1261 gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc
     1321 taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc
     1381 ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc
     1441 tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg
     1501 ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt
     1561 cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg
     1621 agctatgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg
     1681 gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt
     1741 atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag
     1801 gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt
     1861 gctggccttt tgctcacatc ttctttcctg cgttatcccc tgattctgtg gataaccgta
     1921 ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt
     1981 cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc
     2041 cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     2101 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     2161 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     2221 accatgatta cgccaagctt gtaacgactg cagtcttgtt gtcgtttgtc gtcgtcgttc
     2281 tctgtctccc acccaacttt tgccaattgc aacagtgact tctgtacaat ttgcaacaga
     2341 taccaacgtg tcgagtcagc acaaatggcg aggaaggttc gcgtgcattg cttgcgcgcc
     2401 tctttgtcat gggcgcacgc tgatttgtgt gtgatttgca ccaaatttcc tgccgacaca
     2461 acttgggccc gtaccacacg acccgtttct ggacccgttt ctggcccaga aacgcgaatg
     2521 ttgcgaaaga tgcgaccacg tgctcgctgt cgacaacaca aactcccgca caatccccat
     2581 gaaaaaaaaa gtgcttgctg ggcttgtgtc tggcctgtgt gcgtggcgtg gccgcgtgtg
     2641 gcttcgttct tgcttctgct cactcagcct gttacacgca gagtggctgc gagaaacaag
     2701 gagaggggtt tgcggcattg gcgcatgtgc atctgccttt ttcattgagt cagtctgcag
     2761 ctcaagaggt tggtttcgtc gacggatatg agtgagcttc tatggatgat gagaggtggt
     2821 ggtggttgta tggttggtgg ttggtggttg gtgcaaggcg agccacgcga agagcgacaa
     2881 ggatgaggga gaggagagga gactaggaaa gaggtgaagg gaagggaagg gcgagcaatg
     2941 acgacgagat aaaagacagc caccaccgca cccgcacaaa agcattgctc gcacgcatgg
     3001 tcgtgtgtta tgtttcacac caccaacata accaatacaa gccatcacca atacaagcca
     3061 gcgctcaagc acacacgcac gcacgcacac acaatacaca cgcacgtgaa acgttcacat
     3121 tgtccgctca caaatgcctt tcccttctcg tcgctttcct tgcctttttt tcttcccctc
     3181 ctgacttcct tcctgccttc cttccttcct ccccttctcc gtccaccaac accatttatt
     3241 tctctgctct ttttcctcgt gccacgtgag cagtcttttc gcgtcgtctt cttctttgac
     3301 cccgtttacc gtcaacacca tcaacaacca gcaacatgtc tggtacggag tacaagccta
     3361 ccgtccgcct cgctacccgt gacgacgtgc cccgtgctgt ccgtaccctc gccgctgcct
     3421 tcgctgacta ccctgctacc cgtcacaccg tcgaccctga ccgccacatc gagcgtgtca
     3481 ccgagctgca ggagctgttc ctcacccgcg tcggcctcga catcgggaag gtctgggtcg
     3541 ccgacgacgg tgctgctgtc gctgtctgga ccacccctga gtctgtcgag gccggtgctg
     3601 tcttcgctga gatcggcccc cgcatggctg agctgtctgg ctcccgtctc gccgctcagc
     3661 agcagatgga gggcctcctc gccccccacc gccccaagga gcccgcctgg ttcctcgcca
     3721 ccgtcggcgt ctctcctgac caccagggca agggtctcgg ctctgccgtc gtcctccctg
     3781 gcgtcgaggc cgctgagcgt gctggtgtcc ctgctttcct cgagacctcc gctcctcgca
     3841 acctcccctt ctacgagcgt ctcggcttca ccgtcaccgc cgacgtggag gtccccgagg
     3901 gtcctcgtac ctggtgcatg acccgcaagc ccggtggcag cggtgccacc aacttttctc
     3961 tgctcaagca ggctggcgac gttgaagaga accccggacc catgttgagg gtacgaggaa
     4021 gatcgctgga ggcccggttt gtgtacatca tcgtgctggc tgtggtgtac acggccacgt
     4081 acatgtacag cgccacatcc ggtgacccaa gcgtaccggt tggcggcgca gacatgccgc
     4141 cgtggctcgc acagctggga cgaccacgcg cagttgcaga ggagctggca gaacagcaac
     4201 agctgcttga cgcgacgcgg cgcagcgacg acggagccaa cagacacagc aacgatggcg
     4261 ccgccgcagt ctcacaacac cagcagcacc agcagcaggc ggagcacgat ctggccagca
     4321 cacggagtcc aaagacaccg acgcggcaag aggacaaagg cgtggacgag aaggagaaag
     4381 gcatgaagtt cgaggtggag gggggcgaag atggattcgt gcccagggag cacatgaaga
     4441 atgcgatggg tatgccagac actgcagagg ccgacccgtg gaagcggcgc aggcaggaca
     4501 agtggcgcct catcatcttg ttccagcaca acttccccat cctgaagcag gcggtagaag
     4561 ggttccagcg tgccagcgac atcatggtgc ccagcatgat catcgtcgac aactccaagg
     4621 acagggatgc ctcgaacagc gtgtggcttg tggagcgggt ggcagaggtg gtcgtgccag
     4681 accgccagct aaacttccca gagctgcaca actacatggc cacgctggca ctcgagcgcc
     4741 ggctcgaatt ctacttctgg gcccacgcag acaactacgt gctgccgctg gagcctggcc
     4801 gcgacatggg caaggacgcc atcgactgcc tgcgcgagca gatggcgcgg ttccccaact
     4861 ggggcatgat cctcttcgcc tacgaccacc tggccgccta ccgcacgcag acgccagtcc
     4921 aggtgccatg ggacccacac gtgttccagt acggctcgga gtgcgatgcg tatgggcgca
     4981 tccgcgatgc gggctacgac gcgcgcgcgt gcaagattca ctactcgtac gacatgaagc
     5041 gggtgattgg catcctggac acggactcgt acgaccgggt caaggcgaag ctggacgagg
     5101 aggcaaagga caagtccaag cgcaaccagt ggcgcgaggg agccatgtcc gagaaggagc
     5161 agcaatggcg caagcgcatg aaggaggcgt cacgcgcata cttggccgag aagtggggcg
     5221 aggtgaagtg caagctgcgc ggtgtgccgt gccacaagcc ctggccttac tgcccaacgt
     5281 gccccacaga ccttccccag tgctacggca aggaggcgac gtgggagcaa ctcgaccaca
     5341 ttcacgccag ggtgcaccag gtgtttgaca atgatcccga caagccacca ccgctcgagg
     5401 caggcggtag cggtggctcc atggtctcca agggcgagga gaccacgatg ggcgtcatca
     5461 agcccgacat gaagatcaag ctcaagatgg agggcaacgt caacggccac gccttcgtca
     5521 tcgagggcga gggcgagggc aagccctacg acggcaccaa caccatcaac ctcgaggtca
     5581 aggagggcgc ccccctcccc ttctcctacg acatcctcac caccgccttc gcctacggca
     5641 accgcgcctt caccaagtac cccgacgaca tccccaacta cttcaagcag tccttccccg
     5701 agggctactc ctgggagcgc accatgacct tcgaggacaa gggcatcgtc aaggtcaagt
     5761 ccgacatctc tatggaggag gactccttca tctacgagat ccacctcaag ggcgagaact
     5821 tcccccccaa cggccccgtc atgcagaaga agaccaccgg ctgggacgcc tccaccgagc
     5881 gcatgtacgt ccgcgacggc gtcctcaagg gcgacgtgaa gcacaagctc ctcctcgagg
     5941 gcggcggcca ccaccgcgtc gacttcaaga ccatctaccg cgccaagaag gccgtcaagc
     6001 tccccgacta ccacttcgtc gaccaccgca tcgagatcct caaccacaac aaggactaca
     6061 acaaggtcac cgtctacgag tccgccgtcg cccgcaactc caccgacggc atggacgagc
     6121 tgtacaaggc ttaagcgcct cgttggtcat tgccaccttg ttgcctcttt tgctgttgtc
     6181 ctcgtgtcga cgcgtgccac gtgcgcttgt tgtgagttga ttctgcttct tgtgcacgtc
     6241 atgccgagcc cttgggcccc gcaataaacc cactatgcca gtgtaactat ccttcccccc
     6301 aatgtataac tgtgacgatc aatgagcggt gcccgtctag aggatccccg ggtaccgagc
     6361 tcgaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa
     6421 cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc
     6481 accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatggcgcct gatgcggtat
     6541 tttctcctta cgcatctgtg cggtatttca caccgcatat ggtgcactct cagtacaatc
     6601 tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc tgacgcgccc
     6661 tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc
     6721 tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cga
//
