LOCUS       sequence-477931-        7724 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 7724)
  AUTHORS   Drevets DA, Jelinek TA, Freitag NE
  TITLE     Listeria monocytogenes-infected phagocytes can initiate central 
            nervous system infection in mice.
  JOURNAL   Infect Immun. 2001 Mar . 69(3):1344-50.
  PUBMED    11179297
REFERENCE   2  (bases 1 to 7724)
  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: "Infect 
            Immun. 2001 Mar . 69(3):1344-50."
COMMENT     Sequence Label: pNF579
FEATURES             Location/Qualifiers
     source          1..7724
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      72..660
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     561..580
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     814..831
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5264..5285
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5300..5305,5306..5323,5324..5330)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5338..5354
                     /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     5343..5365
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5362..5378
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5362..5378
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5462..5490)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     6085..6105
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(6170..6187)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(6170..6186)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6179..6201)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(6395..6414)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6514..6536
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6574..6592)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6660..6764
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6765..6833,6834..7625)
                     /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(6983..7002)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
       61 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
      121 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
      181 gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta gccgtagtta
      241 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
      301 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
      361 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
      421 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
      481 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
      541 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
      601 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
      661 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
      721 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
      781 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
      841 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctga
      901 tttcactttt tgcattctac aaactgcata actcatatgt aaatcgctcc tttttaggtg
      961 gcacaaatgt gaggcatttt cgctctttcc ggcgaggcta gttaccctta agttattggt
     1021 atgactggtt ttaagcgcaa aaaaagttgc tttttcgtac ctattaatgt atcgttagaa
     1081 aaccgactgt aaaaagtaca gtcggcatta tctcatatta taaaagccag tcattaggcc
     1141 tatctgacaa ttcctgaata gagttcataa acaatcctgc atgataacca tcacaaacag
     1201 aatgatgtac ctgtaaagat agcggtaaat atattgaatt acctttatta atgaattttc
     1261 ctgctgtaat aatgggtaga aggtaattac tattattatt gatatttaag ttaaacccag
     1321 taaatgaagt ccatggaata atagaaagag aaaaagcatt ttcaggtata ggtgttttgg
     1381 gaaacaattt ccccgaacca ttatatttct ctacatcaga aaggtataaa tcataaaact
     1441 ctttgaagtc attctttaca ggagtccaaa taccagagaa tgttttagat acaccatcaa
     1501 aaattgtata aagtggctct aacttatccc aataacctaa ctctccgtcg ctattgtaac
     1561 cagttctaaa agctgtattt gagtttatca cccttgtcac taagaaaata aatgcagggt
     1621 aaaatttata tccttcttgt tttatgtttc ggtataaaac actaatatca atttctgtgg
     1681 ttatactaaa agtcgtttgt tggttcaaat aatgattaaa tatctctttt ctcttccaat
     1741 tgtctaaatc aattttatta aagttcattt gatatgcctc ctaaattttt atctaaagtg
     1801 aatttaggag gcttacttgt ctgctttctt cattagaatc aatccttttt taaaagtcaa
     1861 tattactgta acataaatat atattttaaa aatatcccac tttatccaat tttcgtttgt
     1921 tgaactaatg ggtgctttag ttgaagaata aaagaccaca ttaaaaaatg tggtcttttg
     1981 tgttttttta aaggatttga gcgtagcgaa aaatcctttt ctttcttatc ttgataataa
     2041 gggtaactat tgccgggata gactgtaaca ttctcacgca taaaatcccc tttcattttc
     2101 taatgtaaat ctattacctt attattaatt caattcgctc ataattaatc ctttttctta
     2161 ttacgcaaaa tggcccgatt taagcacacc ctttattccg ttaatgcgcc atgacagcca
     2221 tgataattac taatactagg agaagttaat aaatacgtaa ccaacatgat taacaattat
     2281 tagaggtcat cgttcaaaat ggtatgcgtt ttgacacatc cactatatat ccgtgtcgtt
     2341 ctgtccactc ctgaatccca ttccagaaat tctctagcga ttccagaagt ttctcagagt
     2401 cggaaagttg accagacatt acgaactggc acagatggtc ataacctgaa ggaagatcac
     2461 gataatttcc aagttttaaa caaggatata ttgcagttta aatttcctaa aaaccaatcc
     2521 tataaaatat atggtaatat accttataac ataagtacgg atataatacg caaaattgtt
     2581 tttgatagta tagctaatga gatttattta atcgtggaat acgggtttgc taaaagatta
     2641 ttaaatacaa aacgctcatt ggcattactt ttaatggcag aagttgatat ttctatatta
     2701 agtatggttc caagagaata ttttcatcct aaacctaaag tgaatagctc acttatcaga
     2761 ttaagtagaa aaaaatcaag aatatcacac aaagataaac aaaagtataa ttatttcgtt
     2821 atgaaatggg ttaacaaaga atacaagaaa atatttacaa aaaatcaatt taacaattcc
     2881 ttaaaacatg caggaattga cgatttaaac aatattagct ttgaacaatt cttatctctt
     2941 ttcaatagct ataaattatt taataagtaa gttaagggat gcataaactg catcccttaa
     3001 cttgtttttc gtgtgcctat tttttgtgaa tcgattatgt cttttgcgca gtcggcttaa
     3061 accagttttc gttggtgcga aaaaagagtg tcttgtgaca ctcttaaatt caaaatctat
     3121 cggtcagatt tataccgatt tgattttata tattcttgaa taacatacgc cgagttatca
     3181 cataaaagcg ggaaccaatc atcaaattta aacttcattg cataatccat taaactctta
     3241 aattctacga ttccttgttc atcaataaac tcaatcattt ctttaattaa tttatatcta
     3301 tctgttgttg ttttctttaa taattcatca acatctacac cgccataaac tatcatatct
     3361 tctttttgat atttaaattt attaggatcg tccatgtgaa gcatatatct cacaagacct
     3421 ttcacacttc ctgcaatctg cggaatagtc gcattcaatt cttctgtaat tatttttatc
     3481 tgttcataag atttattacc ctcatacatc actagaatat gataatgctc ttttttcatc
     3541 ctatcttctg tatcagtatc cctatcatgt aatggagaca ctacaaattg aatgtgtaac
     3601 tcttttaaat actctaacca ctcggctttt gctgattctg gatataaaac aaatgtccaa
     3661 ttacgtcctc ttgaattttt cttgttttca gtttctttta ttacattttc gctcatgata
     3721 taataacggt gctaatacat ttaacaaaat ttagtcatag ataggcagca tgccagtgct
     3781 gtctatcttt ttttgtttaa aatgcaccgt attcctcctt tgcatatttt tttattagaa
     3841 taccggttgc atctgatttg ctaatattat atttttcttt gattctattt aatatctcat
     3901 tttcttctgt tgtaagtctt aaagtaacag caactttttt ctcttctttt ctatctacaa
     3961 ccatcactgt acctcccaac atctgttttt ttcactttaa cataaaaaac aaccttttaa
     4021 cattaaaaac ccaatattta tttatttgtt tggacaatgg acaatggaca cctagggggg
     4081 aggtcgtagt acccccctat gttttctccc ctaaataacc ccaaaaatct aagaaaaaaa
     4141 gacctcaaaa aggtctttaa ttaacatctc aaatttcgca tttattccaa tttccttttt
     4201 gcgtgtgatg cgctgcgtcc attaaaaatc ctagagcttt gcaaccgaaa gttaatagct
     4261 gtcgctacta ctttcgctta cgctctaagt atattttaag gactgtcaca cgcaaaaagt
     4321 tttctcggca taaaagtacc tctacatctc taaatcgtct gtacgctgtt tctcacgctt
     4381 tctatcgacc ttctggacat tatcctgtac aacatccata aactgtccca cacgctcgaa
     4441 tttggaatca ttaaagaatt tctctttaag cctattaaac cctttctcaa acccagggaa
     4501 attcgccctc gcagcacgat ataaagtcac tgtactatct tgaaatttct ctgatacatt
     4561 caactgctca ttcaaactat cattctctcg ctttaattta ttaacctctt tacttttttc
     4621 gtgatacccc tctttccatg tattcactac ttctttcaaa ctctctctac gtttttttaa
     4681 ttcttgattt tctgtgtaat agtctgtgct cttaatattt tcgtaatcat caacaatccg
     4741 ttcgaagaga ttgtttcttg caggcgttca aattcatcag cagttaatat ctttctacca
     4801 gtctcttcac gtccagagaa caaacctgta cgctcatttt cataatcaaa gggtttcgta
     4861 gacctcatat gctctattcc actctgtaac tgcttatttg ccttctgtaa ctcatcctta
     4921 acttcttgca gttcctgttt atgaaataca gtatctttct tgtactgatc cagttgctca
     4981 aaaaaatctc ggtcagatgt tactagcaac tcatttacaa gaacagcatc tttcctcgtt
     5041 tttcttgtac ctgttttttg tgattcaata atttctttga cacgttcgtt gtaatcaata
     5101 tttttatcat ttttcaaatc ataattttca cgtgttcgct catggtcaat atcatcattc
     5161 gttctacttt ttcgctctct ttgattatga aattgcatgc cttttagtcc agctggcacg
     5221 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg agttagctca
     5281 ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg tgtggaattg
     5341 tgagcggata acaatttcac acaggaaaca gctatgacat gattacgaat tcgagctcgg
     5401 tacccgggga tcctctagaa ggaggaaaaa tatgagtaaa ggagaagaac ttttcactgg
     5461 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag
     5521 tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac
     5581 tggaaaacta cctgttccat ggccaacact tgtcactact ttctcttatg gtgttcaatg
     5641 cttttcccgt tatccggatc atatgaaacg gcatgacttt ttcaagagtg ccatgcccga
     5701 aggttatgta caggaacgca ctatatcctt caaagatgac gggaactaca agacgcgtgc
     5761 tgaagtcaag tttgaaggtg atacccttgt taatcgtatc gagttaaaag gtattgattt
     5821 taaagaagat ggaaacattc tcggacacaa gctcgagtac aactataact cacacaatgt
     5881 atacatcacg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa ttcgccacaa
     5941 cattgaagat ggatccgttc aactagcaga ccattatcaa caaaatactc caattggcga
     6001 tggccctgtc cttttaccag acaaccatta cctgtcgaca caatctgccc tttcgaaaga
     6061 tcccaacgaa aagcgtgacc acatggtcct tcttgagttt gtaactgctg ctgggattac
     6121 acatggcatg gatgaactat acaaatagct gcaggcatgc aagcttggca ctggccgtcg
     6181 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     6241 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     6301 agttgcgcag cctgaatggc gaatggcgcc tgatgcggta ttttctcctt acgcatctgt
     6361 gcggtatttc acaccgcata tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     6421 taagccagcc ccgacacccg ccaacacccg ctgacgcgcc ctgacgggct tgtctgctcc
     6481 cggcatccgc ttacagacaa gctgtgaccg tctccgggag ctgcatgtgt cagaggtttt
     6541 caccgtcatc accgaaacgc gcgagacgaa agggcctcgt gatacgccta tttttatagg
     6601 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     6661 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     6721 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     6781 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     6841 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     6901 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     6961 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     7021 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     7081 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     7141 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     7201 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     7261 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     7321 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     7381 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     7441 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     7501 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     7561 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     7621 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     7681 aatttaaaag gatctaggtg aagatccttt ttgataatct catg
//
