LOCUS       sequence-10664-e        4299 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4299)
  AUTHORS   Ibarra JA, Knodler LA, Sturdevant DE, Virtaneva K, Carmody AB, 
            Fischer ER, Porcella SF, Steele-Mortimer O
  TITLE     Induction of Salmonella pathogenicity island 1 under different 
            growth conditions can affect Salmonella-host cell interactions in 
            vitro.
  JOURNAL   Microbiology (Reading). 2010 Apr;156(Pt 4):1120-1133. doi: 
            10.1099/mic.0.032896-0. Epub 2009 Dec 24.
  PUBMED    20035008
REFERENCE   2  (bases 1 to 4299)
  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"; journalName: 
            "Microbiology (Reading)."; date: "2010-04"; volume: "156(Pt 
            4):1120-1133. doi"; pages: " 10.1099/mic.0.032896-0. Epub 2009 Dec 
            24"
COMMENT     Sequence Label: pMPMA3dPlac-PfliC-GFPLVA
FEATURES             Location/Qualifiers
     source          1..4299
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     626..645
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     677..693
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     677..693
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     RBS             1069..1080
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1087..1800
                     /codon_start=1
                     /product="S65T variant of Aequorea victoria green 
                     fluorescent protein (Heim et al., 1995)"
                     /label=GFP (S65T)
                     /note="excitable with blue light"
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     CDS             1087..1800
                     /codon_start=1
                     /product="Aequorea victoria green fluorescent protein"
                     /label=GFP
                     /translation="MRKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTFGYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(1117..1145)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1740..1760
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1852..1868)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1853..1869)
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     rep_origin      2154..2699
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     2600..2619
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3311..4102,4103..4171))
                     /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     3934..3953
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4172..4276)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggagctcca
      661 ccgcggtggc ggccgctcta gaactagtgg atcccccggg ctgcaggaat tcgatatcaa
      721 gcttgcatgc ctgcaggtcg actctagaca gtagttaagc gcgttatcgg caatctggag
      781 gcaaagttta atgataattt tgcaaaaata atgcgcggaa taatgatgca taaagcggct
      841 atttcgccgc ctaagaaaaa gatcggggga agtgaaaaat tttctaaagt tcgaaattca
      901 ggtgccgata caagggttac ggtgagaaac cgtgggcaac agcccaataa catcaagttg
      961 taattgataa ggaaaagatc atggcacaag tcattaatac aaacagcctg tcgctgttga
     1021 cccagaataa cctgaacaaa tcccagtccg ctctgggtac ctagaattaa agaggagaaa
     1081 ttaagcatgc gtaaaggaga agaacttttc actggagttg tcccaattct tgttgaatta
     1141 gatggtgatg ttaatgggca caaattttct gtcagtggag agggtgaagg tgatgcaaca
     1201 tacggaaaac ttacccttaa atttatttgc actactggaa aactacctgt tccatggcca
     1261 acacttgtca ctactttcgg ttatggtgtt caatgctttg cgagataccc agatcatatg
     1321 aaacagcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
     1381 tttttcaaag atgacgggaa ctacaagaca cgtgctgaag tcaagtttga aggtgatacc
     1441 cttgttaata gaatcgagtt aaaaggtatt gattttaaag aagatggaaa cattcttgga
     1501 cacaaattgg aatacaacta taactcacac aatgtataca tcatggcaga caaacaaaag
     1561 aatggaatca aagttaactt caaaattaga cacaacattg aagatggaag cgttcaacta
     1621 gcagaccatt atcaacaaaa tactccaatt ggcgatggcc ctgtcctttt accagacaac
     1681 cattacctgt ccacacaatc tgccctttcg aaagatccca acgaaaagag agaccacatg
     1741 gtccttcttg agtttgtaac agctgctggg attacacatg gcatggatga actatacaaa
     1801 aggcctgcag caaacgacga aaactacgct ttagtagctt aataagctta tcgataccgt
     1861 cgacctcgag ggggggcccg gtaccagtcg ggaaacctgt cgtgccagct gcattaatga
     1921 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     1981 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2041 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgac caaaatccct
     2101 taacgtgagt tttcgttcca ctgagcgtca gaccccttaa taagatgatc ttcttgagat
     2161 cgttttggtc tgcgcgtaat ctcttgctct gaaaacgaaa aaaccgcctt gcagggcggt
     2221 ttttcgaagg ttctctgagc taccaactct ttgaaccgag gtaactggct tggaggagcg
     2281 cagtcaccaa aacttgtcct ttcagtttag ccttaaccgg cgcatgactt caagactaac
     2341 tcctctaaat caattaccag tggctgctgc cagtggtgct tttgcatgtc tttccgggtt
     2401 ggactcaaga cgatagttac cggataaggc gcagcggtcg gactgaacgg ggggttcgtg
     2461 catacagtcc agcttggagc gaactgccta cccggaactg agtgtcaggc gtggaatgag
     2521 acaaacgcgg ccataacagc ggaatgacac cggtaaaccg aaaggcagga acaggagagc
     2581 gcacgaggga gccgccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     2641 accactgatt tgagcgtcag atttcgtgat gcttgtcagg ggggcggagc ctatggaaaa
     2701 acggctttgc cgcggccctc tcacttccct gttaagtatc ttcctggcat cttccaggaa
     2761 atctccgccc cgttcgtaag ccatttccgc tcgccgcagt cgaacgaccg agcgtagcga
     2821 gtcagtgagc gaggaagcgg aatatatcct gtatcacata ttctgctgac gcaccggtgc
     2881 agcctttttt ctcctgccac atgaagcctg gtaacaggat tagcagagcg aggtatgtag
     2941 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat
     3001 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     3061 ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc
     3121 gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt
     3181 ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct
     3241 agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt
     3301 ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
     3361 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
     3421 catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat
     3481 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
     3541 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
     3601 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
     3661 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
     3721 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
     3781 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
     3841 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
     3901 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     3961 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     4021 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     4081 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     4141 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     4201 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     4261 aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
