LOCUS       sequence-233745-        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: NK696
FEATURES             Location/Qualifiers
     source          1..6763
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     56..75
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     309..326
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    443..464
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(479..484,485..502,503..509)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    517..533
                     /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     522..544
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     541..557
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     541..557
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2173..2193
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     CDS             2280..2336
                     /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(4704..4721)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4704..4720)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4713..4735)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4929..4948)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5048..5070
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5108..5126)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5194..5298
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5299..5367,5368..6159)
                     /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(5517..5536)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      join(6330..6763,1..155)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
       61 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      121 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      181 ggttcctggc cttttgctgg ccttttgctc acatcttctt tcctgcgtta tcccctgatt
      241 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      301 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      361 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
      421 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
      481 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
      541 caggaaacag ctatgaccat gattacgcca agcttgtaac gactgcagtc ttgttgtcgt
      601 ttgtcgtcgt cgttctctgt ctcccaccca acttttgcca attgcaacag tgacttctgt
      661 acaatttgca acagatacca acgtgtcgag tcagcacaaa tggcgaggaa ggttcgcgtg
      721 cattgcttgc gcgcctcttt gtcatgggcg cacgctgatt tgtgtgtgat ttgcaccaaa
      781 tttcctgccg acacaacttg ggcccgtacc acacgacccg tttctggacc cgtttctggc
      841 ccagaaacgc gaatgttgcg aaagatgcga ccacgtgctc gctgtcgaca acacaaactc
      901 ccgcacaatc cccatgaaaa aaaaagtgct tgctgggctt gtgtctggcc tgtgtgcgtg
      961 gcgtggccgc gtgtggcttc gttcttgctt ctgctcactc agcctgttac acgcagagtg
     1021 gctgcgagaa acaaggagag gggtttgcgg cattggcgca tgtgcatctg cctttttcat
     1081 tgagtcagtc tgcagctcaa gaggttggtt tcgtcgacgg atatgagtga gcttctatgg
     1141 atgatgagag gtggtggtgg ttgtatggtt ggtggttggt ggttggtgca aggcgagcca
     1201 cgcgaagagc gacaaggatg agggagagga gaggagacta ggaaagaggt gaagggaagg
     1261 gaagggcgag caatgacgac gagataaaag acagccacca ccgcacccgc acaaaagcat
     1321 tgctcgcacg catggtcgtg tgttatgttt cacaccacca acataaccaa tacaagccat
     1381 caccaataca agccagcgct caagcacaca cgcacgcacg cacacacaat acacacgcac
     1441 gtgaaacgtt cacattgtcc gctcacaaat gcctttccct tctcgtcgct ttccttgcct
     1501 ttttttcttc ccctcctgac ttccttcctg ccttccttcc ttcctcccct tctccgtcca
     1561 ccaacaccat ttatttctct gctctttttc ctcgtgccac gtgagcagtc ttttcgcgtc
     1621 gtcttcttct ttgaccccgt ttaccgtcaa caccatcaac aaccagcaac atgtctggta
     1681 cggagtacaa gcctaccgtc cgcctcgcta cccgtgacga cgtgccccgt gctgtccgta
     1741 ccctcgccgc tgccttcgct gactaccctg ctacccgtca caccgtcgac cctgaccgcc
     1801 acatcgagcg tgtcaccgag ctgcaggagc tgttcctcac ccgcgtcggc ctcgacatcg
     1861 ggaaggtctg ggtcgccgac gacggtgctg ctgtcgctgt ctggaccacc cctgagtctg
     1921 tcgaggccgg tgctgtcttc gctgagatcg gcccccgcat ggctgagctg tctggctccc
     1981 gtctcgccgc tcagcagcag atggagggcc tcctcgcccc ccaccgcccc aaggagcccg
     2041 cctggttcct cgccaccgtc ggcgtctctc ctgaccacca gggcaagggt ctcggctctg
     2101 ccgtcgtcct ccctggcgtc gaggccgctg agcgtgctgg tgtccctgct ttcctcgaga
     2161 cctccgctcc tcgcaacctc cccttctacg agcgtctcgg cttcaccgtc accgccgacg
     2221 tggaggtccc cgagggtcct cgtacctggt gcatgacccg caagcccggt ggcagcggtg
     2281 ccaccaactt ttctctgctc aagcaggctg gcgacgttga agagaacccc ggacccatgg
     2341 tctccaaggg cgaggagacc acgatgggcg tcatcaagcc cgacatgaag atcaagctca
     2401 agatggaggg caacgtcaac ggccacgcct tcgtcatcga gggcgagggc gagggcaagc
     2461 cctacgacgg caccaacacc atcaacctcg aggtcaagga gggcgccccc ctccccttct
     2521 cctacgacat cctcaccacc gccttcgcct acggcaaccg cgccttcacc aagtaccccg
     2581 acgacatccc caactacttc aagcagtcct tccccgaggg ctactcctgg gagcgcacca
     2641 tgaccttcga ggacaagggc atcgtcaagg tcaagtccga catctctatg gaggaggact
     2701 ccttcatcta cgagatccac ctcaagggcg agaacttccc ccccaacggc cccgtcatgc
     2761 agaagaagac caccggctgg gacgcctcca ccgagcgcat gtacgtccgc gacggcgtcc
     2821 tcaagggcga cgtgaagcac aagctcctcc tcgagggcgg cggccaccac cgcgtcgact
     2881 tcaagaccat ctaccgcgcc aagaaggccg tcaagctccc cgactaccac ttcgtcgacc
     2941 accgcatcga gatcctcaac cacaacaagg actacaacaa ggtcaccgtc tacgagtccg
     3001 ccgtcgcccg caactccacc gacggcatgg acgagctgta caaggctggc ggtagcggtg
     3061 gctccatgtt gagggtacga ggaagatcgc tggaggcccg gtttgtgtac atcatcgtgc
     3121 tggctgtggt gtacacggcc acgtacatgt acagcgccac atccggtgac ccaagcgtac
     3181 cggttggcgg cgcagacatg ccgccgtggc tcgcacagct gggacgacca cgcgcagttg
     3241 cagaggagct ggcagaacag caacagctgc ttgacgcgac gcggcgcagc gacgacggag
     3301 ccaacagaca cagcaacgat ggcgccgccg cagtctcaca acaccagcag caccagcagc
     3361 aggcggagca cgatctggcc agcacacgga gtccaaagac accgacgcgg caagaggaca
     3421 aaggcgtgga cgagaaggag aaaggcatga agttcgaggt ggaggggggc gaagatggat
     3481 tcgtgcccag ggagcacatg aagaatgcga tgggtatgcc agacactgca gaggccgacc
     3541 cgtggaagcg gcgcaggcag gacaagtggc gcctcatcat cttgttccag cacaacttcc
     3601 ccatcctgaa gcaggcggta gaagggttcc agcgtgccag cgacatcatg gtgcccagca
     3661 tgatcatcgt cgacaactcc aaggacaggg atgcctcgaa cagcgtgtgg cttgtggagc
     3721 gggtggcaga ggtggtcgtg ccagaccgcc agctaaactt cccagagctg cacaactaca
     3781 tggccacgct ggcactcgag cgccggctcg aattctactt ctgggcccac gcagacaact
     3841 acgtgctgcc gctggagcct ggccgcgaca tgggcaagga cgccatcgac tgcctgcgcg
     3901 agcagatggc gcggttcccc aactggggca tgatcctctt cgcctacgac cacctggccg
     3961 cctaccgcac gcagacgcta gtccaggtgc catgggaccc acacgtgttc cagtacggct
     4021 cggagtgcga tgcgtatggg cgcatccgcg atgcgggcta cgacgcgcgc gcgtgcaaga
     4081 ttcactactc gtacgacatg aagcgggtga ttggcatcct ggacacggac tcgtacgacc
     4141 gggtcaaggc gaagctggac gaggaggcaa aggacaagtc caagcgcaac cagtggcgcg
     4201 agggagccat gtccgagaag gagcagcaat ggcgcaagcg catgaaggag gcgtcacgcg
     4261 catacttggc cgagaagtgg ggcgaggtga agtgcaagct gcgcggtgtg ccgtgccaca
     4321 agccctggcc ttactgccca acgtgcccca cagaccttcc ccagtgctac ggcaaggagg
     4381 cgacgtggga gcaactcgac cacattcacg ccagggtgca ccaggtgttt gacaatgatc
     4441 ccgacaagcc accaccgctc gaggcataag cgcctcgttg gtcattgcca ccttgttgcc
     4501 tcttttgctg ttgtcctcgt gtcgacgcgt gccacgtgcg cttgttgtga gttgattctg
     4561 cttcttgtgc acgtcatgcc gagcccttgg gccccgcaat aaacccacta tgccagtgta
     4621 actatccttc cccccaatgt ataactgtga cgatcaatga gcggtgcccg tctagaggat
     4681 ccccgggtac cgagctcgaa ttcactggcc gtcgttttac aacgtcgtga ctgggaaaac
     4741 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
     4801 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gcagcctgaa tggcgaatgg
     4861 cgcctgatgc ggtattttct ccttacgcat ctgtgcggta tttcacaccg catatggtgc
     4921 actctcagta caatctgctc tgatgccgca tagttaagcc agccccgaca cccgccaaca
     4981 cccgctgacg cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg
     5041 accgtctccg ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga
     5101 cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat aatggtttct
     5161 tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     5221 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     5281 tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt
     5341 gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct
     5401 gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc
     5461 cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta
     5521 tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac
     5581 tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc
     5641 atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac
     5701 ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg
     5761 gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac
     5821 gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc
     5881 gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt
     5941 gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga
     6001 gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc
     6061 cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag
     6121 atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca
     6181 tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc
     6241 ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca
     6301 gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc
     6361 tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta
     6421 ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt
     6481 ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc
     6541 gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg
     6601 ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg
     6661 tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag
     6721 ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg aca
//
