LOCUS       sequence-324899-        5675 bp    DNA     circular SYN 14-APR-2025
DEFINITION  A plasmid with IFP2.0 coding sequence coded into it. Expresses 
            IFP2.0 with N-term His-tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5675)
  AUTHORS   Chia HE, Zuo T, Koropatkin NM, Marsh ENG, Biteen JS
  TITLE     Imaging living obligate anaerobic bacteria with bilin-binding 
            fluorescent proteins.
  JOURNAL   Curr Res Microb Sci. 2020 Sep;1:1-6. doi: 
            10.1016/j.crmicr.2020.04.001. Epub 2020 May 11.
  PUBMED    33313576
REFERENCE   2  (bases 1 to 5675)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Curr Res 
            Microb Sci."; date: "2020-09"; volume: "1:1-6. doi"; pages: " 
            10.1016/j.crmicr.2020.04.001. Epub 2020 May 11"
COMMENT     Sequence Label: pBAD_IPF2.0
FEATURES             Location/Qualifiers
     source          1..5675
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     118..135
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    150..290
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(216..235)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     376..398
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     536..555
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(735..1613)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        1640..1924
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     1847..1866
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             1951..1973
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             1993..2010
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             2014..2046
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             2059..2079
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             2086..3051
                     /codon_start=1
                     /product="bacteriophytochrome-based monomeric infrared 
                     fluorescent protein (Yu et al., 2014)"
                     /label=IFP2.0
                     /translation="MARDPQPFFPPLYLGGPEITTENCEREPIHIPGSIQPHGALLTAD
                     GHSGEVLQVSLNAATFLGQEPTVLRGQTLAALLPDQWPALQTALPPGCQDALQYRATLD
                     WPAAGHLSLTVHRVAELLILEFEPTEAWDSIGPHALRNAMFALESAPNLRALAEVATQT
                     VRELSGFDRVMLYKFAPDATGEVIAEARREGMQAYLGHRFPASTTPAQARALYTRHLLR
                     LTADTRAAAVPLDPVLNPQTNAPTPLGGAVLRATSPMHMQYLRNMGVGSSLSVSVVVGG
                     QLWGLIVCHHQTPYVLPPDLRTTLEYLGRLLSLQVQRKEA"
     primer_bind     complement(3097..3114)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      3267..3353
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      3445..3472
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        3491..3582
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3583..3651,3652..4443)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3801..3820)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4485..4940
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4572..4591)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4782..4803
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      5051..5639
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5540..5559
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac
       61 cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc
      121 gagtcagtga gcgaggaagc ggaagagcgc ctgatgcggt attttctcct tacgcatctg
      181 tgcggtattt cacaccgcat ctggtgcact ctcagtacaa tctgctctga tgccgcatag
      241 ttaagccagt atacactccg ctatcgctac gtgactgggt catggctgcg ccccgacacc
      301 cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc gcttacagac
      361 aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac
      421 gcgcgaggca gaaggagatg gcgcccaaca gtcccccggc cacggggcct gccaccatac
      481 ccacgccgaa acaagcgctc atgagcccga agtggcgagc ccgatcttcc ccatcggtga
      541 tgtcggcgat ataggcgcca gcaaccgcac ctgtggcgcc ggtgatgccg gccacgatgc
      601 gtccggcgta gaggatctgc tcatgtttga cagcttatca tcgatgcata atgtgcctgt
      661 caaatggacg aagcagggat tctgcaaacc ctatgctact ccgtcaagcc gtcaattgtc
      721 tgattcgtta ccaattatga caacttgacg gctacatcat tcactttttc ttcacaaccg
      781 gcacggaact cgctcgggct ggccccggtg cattttttaa atacccgcga gaaatagagt
      841 tgatcgtcaa aaccaacatt gcgaccgacg gtggcgatag gcatccgggt ggtgctcaaa
      901 agcagcttcg cctggctgat acgttggtcc tcgcgccagc ttaagacgct aatccctaac
      961 tgctggcgga aaagatgtga cagacgcgac ggcgacaagc aaacatgctg tgcgacgctg
     1021 gcgatatcaa aattgctgtc tgccaggtga tcgctgatgt actgacaagc ctcgcgtacc
     1081 cgattatcca tcggtggatg gagcgactcg ttaatcgctt ccatgcgccg cagtaacaat
     1141 tgctcaagca gatttatcgc cagcagctcc gaatagcgcc cttccccttg cccggcgtta
     1201 atgatttgcc caaacaggtc gctgaaatgc ggctggtgcg cttcatccgg gcgaaagaac
     1261 cccgtattgg caaatattga cggccagtta agccattcat gccagtaggc gcgcggacga
     1321 aagtaaaccc actggtgata ccattcgcga gcctccggat gacgaccgta gtgatgaatc
     1381 tctcctggcg ggaacagcaa aatatcaccc ggtcggcaaa caaattctcg tccctgattt
     1441 ttcaccaccc cctgaccgcg aatggtgaga ttgagaatat aacctttcat tcccagcggt
     1501 cggtcgataa aaaaatcgag ataaccgttg gcctcaatcg gcgttaaacc cgccaccaga
     1561 tgggcattaa acgagtatcc cggcagcagg ggatcatttt gcgcttcagc catacttttc
     1621 atactcccgc cattcagaga agaaaccaat tgtccatatt gcatcagaca ttgccgtcac
     1681 tgcgtctttt actggctctt ctcgctaacc aaaccggtaa ccccgcttat taaaagcatt
     1741 ctgtaacaaa gcgggaccaa agccatgaca aaaacgcgta acaaaagtgt ctataatcac
     1801 ggcagaaaag tccacattga ttatttgcac ggcgtcacac tttgctatgc catagcattt
     1861 ttatccataa gattagcgga tactacctga cgctttttat cgcaactctc tactgtttct
     1921 ccatacccgt ttttttgggc tagaaataat tttgtttaac tttaagaagg agatatacat
     1981 atgcggggtt ctcatcatca tcatcatcat ggtatggcta gcatgactgg tggacagcaa
     2041 atgggtcggg atctgtacga gaacctgtac ttccagggct cgagcatggc tcgggaccct
     2101 caacctttct tccctcctct gtatctcgga ggaccagaga tcactactga gaactgcgag
     2161 cgggaaccta tacatatacc aggttctatt caacctcatg gagcactgct cacagccgat
     2221 ggacattctg gcgaggtgct gcaagtttca ctcaacgcag ccactttcct gggccaggaa
     2281 ccaactgttc tgcggggaca gaccctggca gccctgctgc cagaccagtg gcctgctctg
     2341 cagactgccc tgcctccagg ctgccaggac gccctgcagt atcgcgccac cctggattgg
     2401 cccgccgccg gacacctgag cctgaccgtg catagggtgg ccgagctgct gattctggaa
     2461 ttcgagccta ctgaggcatg ggactctatt ggaccccacg ctctgagaaa tgcaatgttc
     2521 gctctggaga gtgctcctaa tctccgggca ctggctgaag tcgcaaccca aacagtccgg
     2581 gaactgtcag gtttcgaccg ggtgatgctg tacaagtttg caccagacgc aacaggagag
     2641 gttattgccg aagcaaggcg cgagggcatg caggcttacc tcgggcatag gtttcccgca
     2701 tccaccaccc ctgcacaagc tagggccctc tacacaagac acctgctccg gctgaccgca
     2761 gacaccaggg ctgcagcagt gcccctcgac cccgtgctga atccccagac aaatgctcct
     2821 acacctctgg gcggagctgt cctcagagct acatccccaa tgcacatgca gtacctgagg
     2881 aatatgggag tgggctcctc cctgagcgtc agcgtcgtgg tcggcggcca gctgtgggga
     2941 ctgattgtct gccaccatca gacaccctac gtgctgccac ccgatctgcg gaccaccctg
     3001 gagtatctgg ggaggctcct gtccctgcag gtgcagagga aagaagcctg agaattcgaa
     3061 gcttggctgt tttggcggat gagagaagat tttcagcctg atacagatta aatcagaacg
     3121 cagaagcggt ctgataaaac agaatttgcc tggcggcagt agcgcggtgg tcccacctga
     3181 ccccatgccg aactcagaag tgaaacgccg tagcgccgat ggtagtgtgg ggtctcccca
     3241 tgcgagagta gggaactgcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3301 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgg
     3361 gagcggattt gaacgttgcg aagcaacggc ccggagggtg gcgggcagga cgcccgccat
     3421 aaactgccag gcatcaaatt aagcagaagg ccatcctgac ggatggcctt tttgcgtttc
     3481 tacaaactct tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     3541 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     3601 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     3661 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     3721 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     3781 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga cgccgggcaa
     3841 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     3901 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     3961 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     4021 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     4081 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgcagc aatggcaaca
     4141 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     4201 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     4261 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     4321 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     4381 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     4441 taactgtcag accaagttta ctcatatata ctttagattg atttacgcgc cctgtagcgg
     4501 cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc
     4561 cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc
     4621 ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct
     4681 cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac
     4741 ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac
     4801 tggaacaaca ctcaacccta tctcgggcta ttcttttgat ttataaggga ttttgccgat
     4861 ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa
     4921 aatattaacg tttacaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4981 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     5041 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     5101 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     5161 taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag
     5221 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     5281 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     5341 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     5401 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     5461 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     5521 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     5581 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     5641 acgccagcaa cgcggccttt ttacggttcc tggcc
//
