LOCUS       sequence-383991-        4656 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  A strong P7 promoter flanked by Bxb1 recombination sites attB and 
            attP. Downstream of attP is a strong RBS and YFP coding sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4656)
  AUTHORS   Pandey A, Rodriguez ML, Poole W, Murray RM
  TITLE     Characterization of Integrase and Excisionase Activity in a 
            Cell-Free Protein Expression System Using a Modeling and Analysis 
            Pipeline.
  JOURNAL   ACS Synth Biol. 2023 Feb 17;12(2):511-523. doi: 
            10.1021/acssynbio.2c00534. Epub 2023 Jan 30.
  PUBMED    36715625
REFERENCE   2  (bases 1 to 4656)
  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"; doi: 
            "10.1021/acssynbio.2c00534"; journalName: "ACS Synth Biol"; date: 
            "2023-02-17- 17"; volume: "12"; issue: "2"; pages: "511-523"
COMMENT     Sequence Label: attB-P7-attP-YFP
FEATURES             Location/Qualifiers
     source          1..4656
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(100..129)
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     protein_bind    181..230
                     /label=Bxb1 attB
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="attB site for the Mycopbacteriophage Bxb1 integrase 
                     (Xu et al., 2013)"
     protein_bind    285..332
                     /label=Bxb1 attP
                     /bound_moiety="Mycobacteriophage Bxb1 integrase"
                     /note="Minimal attP site for the Mycopbacteriophage Bxb1 
                     integrase (Voutev and Mann, 2017)"
     misc_RNA        339..388
                     /label=sTRSV HHRz
                     /note="hammerhead ribozyme from the tobacco ringspot virus 
                     satellite RNA (Khvorova et al., 2003)"
     RBS             482..490
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     CDS             503..568
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     terminator      1338..1409
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1425..1452
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     misc_feature    1459..1479
                     /label=BioBrick suffix
                     /note="universal suffix for all parts"
     terminator      1515..1546
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     terminator      1671..1765
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     rep_origin      2257..2479
                     /label=pSC101 ori
                     /note="low-copy replication origin that requires the Rep101
                     protein"
     CDS             2527..3477
                     /codon_start=1
                     /gene="rep101"
                     /product="RepA protein needed for replication with the 
                     pSC101 origin"
                     /label=Rep101
                     /translation="MSELVVFKANELAISRYDLTEHETKLILCCVALLNPTIENPTRKE
                     RTVSFTYNQYAQMMNISRENAYGVLAKATRELMTRTVEIRNPLVKGFEIFQWTNCAKFS
                     SEKLELVFSEGILPYLFQLKKFIKYNLEHVKSFENKYSMRIYEWLLKELTQKKTHKANI
                     EISLDEFKFMLMLENNYHEFKRLNQWVLKPISKDLNTYSNMKLVVDKRGRPTDTLIFQV
                     ELDRQMDLVTELENNQIKMNGDKIPTTITSDSYLHNGLRKTLHDALTAKIQFTSFEAKF
                     LGDMQSKYDLNGSFSWLTQKQRTTLENILAKYGRI"
     CDS             join(3766..3834,3835..4626)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITTSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ggtgtcacca ccctgccctt tttctttaaa accgaaaaga ttacttcgcg tttgccacct
       61 gacgtctaag aaaaggaata ttcagcaatt tgcccgtgcc gaagaaaggc ccacccgtga
      121 aggtgagcca gtgagttgat tgctacgtaa ttagttagtt agcccttagt gactcgaatt
      181 tcggccggct tgtcgacgac ggcggtctcc gtcgtcagga tcatccgggc aagctttcca
      241 cacattatag gtacaaaaag atgcgaaagc aaataaattt tttgggtttg taccgtacac
      301 cactgagacc gcggtggttg accagacaaa ccacgaagct gtcaccggat gtgctttccg
      361 gtctgatgag tccgtgagga cgaaacagcc tctacaaata attttgttta agggcccaag
      421 ttcacttaaa aaggagatca acaatgaaag caattttcgt actgaaacat cttaatcatg
      481 ctaaggaggt tttctaatga tggactacaa ggaccatgac ggtgattaca aggatcatga
      541 catcgactac aaggatgacg atgacaaggc aactagcggc atggttagta aaggagaaga
      601 acttttcact ggagttgtcc caattttagt tgaactagat ggcgacgtga acggtcataa
      661 gttcagtgtc tccggcgaag gtgagggtga tgcaacgtac ggtaagttaa ctttgaagtt
      721 aatatgtaca accggcaagc tgcctgttcc ctggcctacc ctggtgacaa cgttaggtta
      781 tgggttgatg tgctttgcta gatacccaga tcacatgaaa aggcatgact tctttaaatc
      841 tgcaatgcca gaaggttacg tccaagaacg tactattttc tttaaagatg acggtaatta
      901 taaaactagg gctgaagtta aattcgaagg tgacacactt gtaaatcgaa tagagttaaa
      961 ggggattgat ttcaaagagg atggtaatat tctaggccat aaacttgaat ataactataa
     1021 ttcacacaac gtttacatta ccgccgacaa gcagaagaat ggaatcaaag ccaattttaa
     1081 gattagacac aatattgagg atggtggagt acagcttgct gatcattacc aacaaaatac
     1141 cccgatcggt gatggaccag ttttgctacc cgataaccat tatctgtcct atcaaagcaa
     1201 attgtcaaaa gatcctaacg aaaaaagaga ccacatggta ctcttggaat ttgtaacagc
     1261 tgctgggatt acacatggca tggatgaact atacaaaggt tctggtaccg cagcatgatg
     1321 atactagagc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcccttcgtt
     1381 ttatctgttg tttgtcggtg aacgctctct actagagtca cactggctca ccttcgggtg
     1441 ggcctttctg cgtttatata ctagtagcgg ccgctgcagg agtcactaag ggttagttag
     1501 ttagattagc agaaagtcaa aagcctccga ccggaggctt ttgactaaaa cttcccttgg
     1561 ggttatcatt ggggctcact caaaggcggt aatcagataa aaaaaatcct tagctttcgc
     1621 tgaggatgat ttctgctaga gctgtcagac caagtttacg agctcgcttg gactcctgtt
     1681 gatagatcca gtaatgacct cagaactcca tctggatttg ttcagaacgc tcggttgccg
     1741 ccgggcgttt tttattggtg agaatccaag cactagggac agtaagacgg gtaagcctgt
     1801 tgatgatacc gctgccttac tgggtgcatt agccagtctg aatgacctgt cacggggtaa
     1861 tccgaagtgg ccagactgga aaatcagagg gcaggaactg ctgaacagca aaaagtcaga
     1921 tagcaccgca tagcagaccc gccataaaac gccctgagaa gcccgtgacg ggcttttctt
     1981 gtattatggg tagtttcctt gcatgaatcc ataaaaggcg cctgtagtgc catttacccc
     2041 cattcactgc cagagccgtg agtgcagcga actgaatgtc acgaaaaaga cggcgactca
     2101 ggtgcctgat ggtcggagac aaaaggaata ttcagcgatt tgcccgagct tgcgagggtg
     2161 ctacttaagc ctttagggtt ttaaggtctg ttttgtagag gagcaaacag cgtttgcgac
     2221 atccttttgt aatactgcgg aactgactaa agtagtgagt tatacacagg gctgggatct
     2281 attcttttta tcttttttta ttctttcttt attctataaa ttataaccac ttgaatataa
     2341 acaaaaaaaa cacacaaagg tctagcggaa tttacagagg gtctagcaga atttacaagt
     2401 tttccagcaa aggtctagca gaatttacag atacccacaa ctcaaaggaa aaggacatgt
     2461 aattatcatt gactagccca tctcaattgg tatagtgatt aaaatcacct agaccaattg
     2521 agatgtatgt ctgaattagt tgttttcaaa gcaaatgaac tagcgattag tcgctatgac
     2581 ttaacggagc atgaaaccaa gctaatttta tgctgtgtgg cactactcaa ccccacgatt
     2641 gaaaacccta caaggaaaga acggacggta tcgttcactt ataaccaata cgctcagatg
     2701 atgaacatca gtagggaaaa tgcttatggt gtattagcta aagcaaccag agagctgatg
     2761 acgagaactg tggaaatcag gaatcctttg gttaaaggct ttgagatttt ccagtggaca
     2821 aactgtgcca agttctcaag cgaaaaatta gaattagttt ttagtgaagg gatattgcct
     2881 tatcttttcc agttaaaaaa attcataaaa tataatctgg aacatgttaa gtcttttgaa
     2941 aacaaatact ctatgaggat ttatgagtgg ttattaaaag aactaacaca aaagaaaact
     3001 cacaaggcaa atatagagat tagccttgat gaatttaagt tcatgttaat gcttgaaaat
     3061 aactaccatg agtttaaaag gcttaaccaa tgggttttga aaccaataag taaagatttg
     3121 aacacttaca gcaatatgaa attggtggtt gataagcgag gccgcccgac tgatacgttg
     3181 attttccaag ttgaactaga tagacaaatg gatctcgtaa ccgaacttga gaacaaccag
     3241 ataaaaatga atggtgacaa aataccaaca accattacat cagattccta cctacataac
     3301 ggactaagaa aaacactaca cgatgcttta actgcaaaaa ttcagttcac cagttttgag
     3361 gcaaaatttt tgggtgacat gcaaagtaag tatgatctca atggttcgtt ctcatggctc
     3421 acgcaaaaac aacgaaccac actagagaac atactggcta aatacggaag gatctgaggt
     3481 tcttatggct cttgtatcta tcagtgaagc atcaagacta acaaacaaaa gtagaacaac
     3541 tgttcaccgt tacatatcaa agggaaaact gtccatatgc acagatgaaa acggtgtaaa
     3601 aaagatagat acatcagagc ttttacgagt ttttggtgca ttcaaagctg ttcaccatga
     3661 acagatcgac aatgtaacta ctagagctaa atacattcaa atatctatcc gctcatgaga
     3721 caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag tattcaacat
     3781 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     3841 gaaacgctgg tgaaagtaaa agatgccgaa gatcagttgg gcgcacgtgt gggttacatc
     3901 gaactggacc tcaacagcgg taagattctt gagagttttc gccccgaaga acgtttccca
     3961 atgatgagca cttttaaagt tctgctctgt ggcgcggtat tatcccgtat tgacgccggg
     4021 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     4081 gtcacagaaa agcatcttac ggacggcatg acagtacgcg aattatgcag cgctgccata
     4141 accacgagtg ataacacggc ggccaactta cttctgacaa cgatcggggg accgaaggag
     4201 cttaccgctt ttttgcacaa catgggtgat catgtaactc gccttgatcg ttgggaaccg
     4261 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agctatggca
     4321 acaacgttgc gcaaactctt aactggcgaa cttcttactc tcgcttcccg gcaacaatta
     4381 atagactgga tggaggcgga taaagttgcg ggaccacttc tgcgctcggc ccttccggct
     4441 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtcccgcgg tattattgca
     4501 gccctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagccag
     4561 gcaactatgg acgaacgtaa tcgccagatc gctgagatag gtgcctccct gattaagcat
     4621 tggtaataat actagctccg gcaaaaaaac gggcaa
//
