LOCUS       sequence-233701-        6921 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 6921)
  TITLE     King Lab plasmids
REFERENCE   2  (bases 1 to 6921)
  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: NK691
FEATURES             Location/Qualifiers
     source          1..6921
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      64..519
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(151..170)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     361..382
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        545..649
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(650..718,719..1510)
                     /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(868..887)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1681..2269
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2170..2189
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2423..2440
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2557..2578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2593..2598,2599..2616,2617..2623)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2631..2647
                     /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     2636..2658
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2655..2671
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2655..2671
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     2690..2710
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2692..2710
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     repeat_region   2734..2796
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     repeat_region   6745..6779
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(6798..6817)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6799..6817)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6827..6844)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6827..6843)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6836..6858)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
ORIGIN
        1 atagcgaaga ggcccgcacc gatcgccctt cccaacagtt gcgcagcctg aatggcgaat
       61 gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga
      121 ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg
      181 ccacgttcgc cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat
      241 ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg
      301 ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata
      361 gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt
      421 tataagggat tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat
      481 ttaacgcgaa ttttaacaaa atattaacgc ttacaattta ggtggcactt ttcggggaaa
      541 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat
      601 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca
      661 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca
      721 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta
      781 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt
      841 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc
      901 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc
      961 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc
     1021 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa
     1081 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga
     1141 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat
     1201 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca
     1261 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc
     1321 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat
     1381 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag
     1441 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa
     1501 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca
     1561 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     1621 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     1681 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     1741 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     1801 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     1861 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     1921 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     1981 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     2041 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     2101 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     2161 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     2221 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     2281 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     2341 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
     2401 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
     2461 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
     2521 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
     2581 ggcaccccag gctttacact ttatgctccc ggctcgtatg ttgtgtggaa ttgtgagcgg
     2641 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     2701 tcactaaagg gaacaaaagc tgggtactac tagccctaga aagataatca tattgtgacg
     2761 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
     2821 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     2881 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     2941 aaatttcttc tataaagtaa caaaactttt agctagtggt acaagatcta agcttgtaac
     3001 gactgcagtc ttgttgtcgt ttgtcgtcgt cgttctctgt ctcccaccca acttttgcca
     3061 attgcaacag tgacttctgt acaatttgca acagatacca acgtgtcgag tcagcacaaa
     3121 tggcgaggaa ggttcgcgtg cattgcttgc gcgcctcttt gtcatgggcg cacgctgatt
     3181 tgtgtgtgat ttgcaccaaa tttcctgccg acacaacttg ggcccgtacc acacgacccg
     3241 tttctggacc cgtttctggc ccagaaacgc gaatgttgcg aaagatgcga ccacgtgctc
     3301 gctgtcgaca acacaaactc ccgcacaatc cccatgaaaa aaaaagtgct tgctgggctt
     3361 gtgtctggcc tgtgtgcgtg gcgtggccgc gtgtggcttc gttcttgctt ctgctcactc
     3421 agcctgttac acgcagagtg gctgcgagaa acaaggagag gggtttgcgg cattggcgca
     3481 tgtgcatctg cctttttcat tgagtcagtc tgcagctcaa gaggttggtt tcgtcgacgg
     3541 atatgagtga gcttctatgg atgatgagag gtggtggtgg ttgtatggtt ggtggttggt
     3601 ggttggtgca aggcgagcca cgcgaagagc gacaaggatg agggagagga gaggagacta
     3661 ggaaagaggt gaagggaagg gaagggcgag caatgacgac gagataaaag acagccacca
     3721 ccgcacccgc acaaaagcat tgctcgcacg catggtcgtg tgttatgttt cacaccacca
     3781 acataaccaa tacaagccat caccaataca agccagcgct caagcacaca cgcacgcacg
     3841 cacacacaat acacacgcac gtgaaacgtt cacattgtcc gctcacaaat gcctttccct
     3901 tctcgtcgct ttccttgcct ttttttcttc ccctcctgac ttccttcctg ccttccttcc
     3961 ttcctcccct tctccgtcca ccaacaccat ttatttctct gctctttttc ctcgtgccac
     4021 gtgagcagtc ttttcgcgtc gtcttcttct ttgaccccgt ttaccgtcaa caccatcaac
     4081 aaccagcaac atgttgaggg tacgaggaag atcgctggag gcccggtttg tgtacatcat
     4141 cgtgctggct gtggtgtaca cggccacgta catgtacagc gccacatccg gtgacccaag
     4201 cgtaccggtt ggcggcgcag acatgccgcc gtggctcgca cagctgggac gaccacgcgc
     4261 agttgcagag gagctggcag aacagcaaca gctgcttgac gcgacgcggc gcagcgacga
     4321 cggagccaac agacacagca acgatggcgc cgccgcagtc tcacaacacc agcagcacca
     4381 gcagcaggcg gagcacgatc tggccagcac acggagtcca aagacaccga cgcggcaaga
     4441 ggacaaaggc gtggacgaga aggagaaagg catgaagttc gaggtggagg ggggcgaaga
     4501 tggattcgtg cccagggagc acatgaagaa tgcgatgggt atgccagaca ctgcagaggc
     4561 cgacccgtgg aagcggcgca ggcaggacaa gtggcgcctc atcatcttgt tccagcacaa
     4621 cttccccatc ctgaagcagg cggtagaagg gttccagcgt gccagcgaca tcatggtgcc
     4681 cagcatgatc atcgtcgaca actccaagga cagggatgcc tcgaacagcg tgtggcttgt
     4741 ggagcgggtg gcagaggtgg tcgtgccaga ccgccagcta aacttcccag agctgcacaa
     4801 ctacatggcc acgctggcac tcgagcgccg gctcgaattc tacttctggg cccacgcaga
     4861 caactacgtg ctgccgctgg agcctggccg cgacatgggc aaggacgcca tcgactgcct
     4921 gcgcgagcag atggcgcggt tccccaactg gggcatgatc ctcttcgcct acgaccacct
     4981 ggccgcctac cgcacgcaga cgccagtcca ggtgccatgg gacccacacg tgttccagta
     5041 cggctcggag tgcgatgcgt atgggcgcat ccgcgatgcg ggctacgacg cgcgcgcgtg
     5101 caagattcac tactcgtacg acatgaagcg ggtgattggc atcctggaca cggactcgta
     5161 cgaccgggtc aaggcgaagc tggacgagga ggcaaaggac aagtccaagc gcaaccagtg
     5221 gcgcgaggga gccatgtccg agaaggagca gcaatggcgc aagcgcatga aggaggcgtc
     5281 acgcgcatac ttggccgaga agtggggcga ggtgaagtgc aagctgcgcg gtgtgccgtg
     5341 ccacaagccc tggccttact gcccaacgtg ccccacagac cttccccagt gctacggcaa
     5401 ggaggcgacg tgggagcaac tcgaccacat tcacgccagg gtgcaccagg tgtttgacaa
     5461 tgatcccgac aagccaccac cgctcgaggc aggcggtagc ggtggctcca tggtctccaa
     5521 gggcgaggag accacgatgg gcgtcatcaa gcccgacatg aagatcaagc tcaagatgga
     5581 gggcaacgtc aacggccacg ccttcgtcat cgagggcgag ggcgagggca agccctacga
     5641 cggcaccaac accatcaacc ttgaggtcaa agaaggtgct cctctgccct ttagctacga
     5701 catcctcacc accgccttct cctacggcaa ccgcgccttc accaagtacc ccgacgacat
     5761 ccccaactac ttcaagcagt ccttccccga gggctactcc tgggagcgca ccatgacctt
     5821 cgaggacaag ggcatcgtca aggtcaagtc cgacatctct atggaggagg actccttcat
     5881 ctacgagatc cacctcaagg gcgagaactt ccctcccaac ggccccgtca tgcagaagga
     5941 gactacgggt tgggacgcct ccaccgagcg catgtacgtc cgcgacggtg ttctcaaggg
     6001 cgacgtgaag atgaaacttc tccttgaggg cggcggccac caccgcgtcg acttcaagac
     6061 catctaccgc gccaagaagg ccgtcaagct ccccgactac cacttcgtcg accaccgcat
     6121 cgagatcctc aaccacgaca aggactacaa caaagtcact gtctacgaga tcgccgtcgc
     6181 ccgcaactct accgacggca tggacgagct ttacaaggct taagcgcctc gttggtcatt
     6241 gccaccttgt tgcctctttt gctgttgtcc tcgtgtcgac gcgtgccacg tgcgcttgtt
     6301 gtgagttgat tctgcttctt gtgcacgtca tgccgagccc ttgggccccg caataaaccc
     6361 actatgccag tgtaactatc cttcccccca atgtataact gtgacgatca atgagcggtg
     6421 cccgagatct cctgcagggg taccgctagc tctagcacta gtggccggcc gatatctata
     6481 acaagaaaat atatatataa taagttatca cgtaagtaga acatgaaata acaatataat
     6541 tatcgtatga gttaaatctt aaaagtcacg taaaagataa tcatgcgtca ttttgactca
     6601 cgcggtcgtt atagttcaaa atcagtgaca cttaccgcat tgacaagcac gcctcacggg
     6661 agctccaagc ggcgactgag atgtcctaaa tgcacagcga cggattcgcg ctatttagaa
     6721 agagagagca atatttcaag aatgcatgcg tcaattttac gcagactatc tttctagggc
     6781 tagatagctc caattcgccc tatagtgagt cgtattacgc gcgctcactg gccgtcgttt
     6841 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     6901 cccctttcgc cagctggcgt a
//
