LOCUS       sequence-203076-        3653 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 3653)
  TITLE     Ristow lab plasmids
REFERENCE   2  (bases 1 to 3653)
  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: pBlue-hFX-HA(NcoI/EcoRI)
FEATURES             Location/Qualifiers
     source          1..3653
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     44..60
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     45..61
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             726..752
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(786..802)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(786..802)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     primer_bind     complement(839..859)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(839..857)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(878..894)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(878..894)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(891..913)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    902..918
                     /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)."
     promoter        complement(join(926..932,933..950,951..956))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    971..992
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1109..1126)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1280..1868)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1360..1379)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2039..2830,2831..2899))
                     /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     2662..2681
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2900..3004)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      3031..3487
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3118..3137)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3328..3349
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3613..3635
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3627..3644
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3628..3644
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 taatacgact cactataggg cgaattgggt accgggcccc ccctcgaggt cgacggtatc
       61 gataagcttg atatcgaatt cctgcagccc ccatgggatg gactctcggg cgccgcgcag
      121 tagccggcct cctggcgtca cccagcccgg cccaggccca gaccctcacc cgggtcccgc
      181 ggccggcaga gttggcccca ctctgcggcc gccgtggcct gcgcaccgac atcgatgcga
      241 cctgcacgcc ccgccgcgca agttcgaacc aacgtggcct caaccagatt tggaatgtca
      301 aaaagcagag tgtctatttg atgaatttga ggaaatctgg aactttgggc cacccaggct
      361 ctctagatga gaccacctat gaaagactag cagaggaaac gctggactct ttagcagagt
      421 tttttgaaga ccttgcagac aagccataca cgtttgagga ctatgatgtc tcctttggga
      481 gtggtgtctt aactgtcaaa ctgggtggag atctaggaac ctatgtgatc aacaagcaga
      541 cgccaaacaa gcaaatctgg ctatcttctc catccagtgg acctaagcgt tatgactgga
      601 ctgggaaaaa ctgggtgtac tcccacgacg gcgtgtccct ccatgagctg ctggccgcag
      661 agctcactaa agccttaaaa accaaactgg acttgtcttc cttggcctat tccggaaaag
      721 atgcttaccc ttacgacgta ccagactacg cagacacgta ccgatacatc tagaattccc
      781 agggggatcc actagttcta gagcggccgc caccgcggtg gagctccagc ttttgttccc
      841 tttagtgagg gttaattgcg cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa
      901 attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct
      961 ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc
     1021 agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg
     1081 gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc
     1141 ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag
     1201 gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa
     1261 aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc
     1321 gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc
     1381 ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg
     1441 cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt
     1501 cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc
     1561 gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc
     1621 cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag
     1681 agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg
     1741 ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa
     1801 ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag
     1861 gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact
     1921 cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa
     1981 attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt
     2041 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
     2101 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
     2161 gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc
     2221 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
     2281 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
     2341 ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
     2401 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
     2461 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
     2521 tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
     2581 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
     2641 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
     2701 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
     2761 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
     2821 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
     2881 ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt
     2941 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc
     3001 cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca ttaagcgcgg
     3061 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     3121 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     3181 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     3241 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     3301 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     3361 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     3421 taaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgctt
     3481 acaatttcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     3541 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     3601 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgagcgc gcg
//
