LOCUS       sequence-60294-e        5601 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 5601)
  AUTHORS   Piatkevich KD, Subach FV, Verkhusha VV
  TITLE     Far-red light photoactivatable near-infrared fluorescent proteins 
            engineered from a bacterial phytochrome.
  JOURNAL   Nat Commun. 2013 Jul 10;4:2153. doi: 10.1038/ncomms3153.
  PUBMED    23842578
REFERENCE   2  (bases 1 to 5601)
  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: "Nat 
            Commun."; date: "2013-07-10"; volume: "4:2153. doi"; pages: " 
            10.1038/ncomms3153"
COMMENT     Sequence Label: pBAD/HisB-PAiRFP2
FEATURES             Location/Qualifiers
     source          1..5601
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..285
                     /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     208..227
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     CDS             331..348
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             352..384
                     /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             join(388..396,397..411)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     primer_bind     complement(2002..2019)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      2172..2258
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2350..2377
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        2396..2487
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2488..2556,2557..3348)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2706..2725)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3519..4107
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4008..4027
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4261..4278
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    4293..4433
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(4359..4378)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4519..4541
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4697..5575)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
ORIGIN
        1 aagaaaccaa ttgtccatat tgcatcagac attgccgtca ctgcgtcttt tactggctct
       61 tctcgctaac caaaccggta accccgctta ttaaaagcat tctgtaacaa agcgggacca
      121 aagccatgac aaaaacgcgt aacaaaagtg tctataatca cggcagaaaa gtccacattg
      181 attatttgca cggcgtcaca ctttgctatg ccatagcatt tttatccata agattagcgg
      241 atcctacctg acgcttttta tcgcaactct ctactgtttc tccatacccg ttttttgggc
      301 taacaggagg aattaaccat ggggggttct catcatcatc atcatcatgg tatggctagc
      361 atgactggtg gacagcaaat gggtcgggat ctgtacgacg atgacgataa ggatccgagc
      421 tcgagatcta tggcatccac ggactatcat gtcgacctga ccaattgcga ccgcgaaccc
      481 attcatattc caggatatat tcaaccgcat ggctgcctca tcgcctgcga taacgccatg
      541 cgaatggtgc tgcggcattc cgaaaactgc ggcgaactgc tgggacttga gggcgatctc
      601 aacggcagga ccgccgagga tgtgctcggc aaaaggctcg tccatgatct ccgcaacgcg
      661 ctcaccgtca ccggtaaaac cacgcgcccc gccatgctgc ccgcaatgga aacaagcgac
      721 ggccgcagct tcgatatttc actccatcgc tacaaatcca ccaccatcat cgaattcgag
      781 ccctcggaca gcgacacgca gccgctcggc acggcgcgca agatggtgga ccgcatccgc
      841 gaagccgaca gcgtcgaaag cctgatctcc agaaccacac gcctggtaaa ggcgacgctg
      901 ggttacgacc gtgtgctgat ctaccgcttc gaggaagacg gcgccggcaa ggtcgtgtcg
      961 gaggccaagc agcccgagct ggagagcttt ctcgggcaat atttccccgc cagcgatatt
     1021 ccacagcagg ctcgcgcact ttatctcaag aacaccttgc gcatcatctc cgatgccagc
     1081 ggcaccccca ttccggtcct gccagccgtc gatgtctccg gtgagccgct cgatctctct
     1141 tacgcgcatt tgcgcaactt ctcgcccatc cattgcgaat atctccgcaa tatgggtgtt
     1201 gccgcctcca tgtcgatttc tgtgattgtc gatgacgccc tgtggggcct gatcgtctgc
     1261 caccattgtt cgccgcgtgt gctgagcatg ccagttcgga tcgctgcggc gatgttcggg
     1321 gaatttttct ccatgttcct gcaggttctg aaacagaagc gccgcctcga caccatcaac
     1381 cacgcccatg cggcgctcga ccggtttctg cggctggccg cccatcgggc caacctcgaa
     1441 gaactgctgg tcgacagttt tcaggatttt gcccgcctga taccctgcga cggtgtcggg
     1501 ctgtgggtcg gcaacaactg gcacggccat ggcgcaacgc caccgcatga tgccattcca
     1561 aggctggcac gtttcgtcgc ttcggtttcg gaaggcaggg tctgggcaac acacgccctg
     1621 tcgcaggcca taccggaagc ggaaatatac gccgacaccg cggccggaat gcttgctatc
     1681 ccgatttcgc aggtcaagag cgactatctc ctgtttttcc gcaaggagat cgtgcagaac
     1741 ctgaactggg ccggcaatcc ggaaaaatcc tatgaaaccg gcccgatggg cgaccgcctc
     1801 acgccgcgta aaagcttcgc aatctggaaa gagtccgttc gcctgcaggc acagccctgg
     1861 tcggaagccg accgggaaat cgccgaaacc gcgagaatcg cgcttgtcgg agtggcgttc
     1921 caccacagcg agttgatggc aggcctcgag tacaagtaat tcgaagcttg gctgttttgg
     1981 cggatgagag aagattttca gcctgataca gattaaatca gaacgcagaa gcggtctgat
     2041 aaaacagaat ttgcctggcg gcagtagcgc ggtggtccca cctgacccca tgccgaactc
     2101 agaagtgaaa cgccgtagcg ccgatggtag tgtggggtct ccccatgcga gagtagggaa
     2161 ctgccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt cgttttatct
     2221 gttgtttgtc ggtgaacgct ctcctgagta ggacaaatcc gccgggagcg gatttgaacg
     2281 ttgcgaagca acggcccgga gggtggcggg caggacgccc gccataaact gccaggcatc
     2341 aaattaagca gaaggccatc ctgacggatg gcctttttgc gtttctacaa actcttttgt
     2401 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     2461 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     2521 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     2581 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     2641 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     2701 gttctgctat gtggcgcggt attatcccgt gttgacgccg ggcaagagca actcggtcgc
     2761 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     2821 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     2881 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     2941 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     3001 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     3061 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     3121 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     3181 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     3241 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     3301 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     3361 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     3421 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     3481 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     3541 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     3601 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     3661 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     3721 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     3781 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     3841 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     3901 cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     3961 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     4021 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     4081 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     4141 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     4201 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     4261 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     4321 ctgtgcggta tttcacaccg catatggtgc actctcagta caatctgctc tgatgccgca
     4381 tagttaagcc agtatacact ccgctatcgc tacgtgactg ggtcatggct gcgccccgac
     4441 acccgccaac acccgctgac gcgccctgac gggcttgtct gctcccggca tccgcttaca
     4501 gacaagctgt gaccgtctcc gggagctgca tgtgtcagag gttttcaccg tcatcaccga
     4561 aacgcgcgag gcagcagatc aattcgcgcg cgaaggcgaa gcggcatgca taatgtgcct
     4621 gtcaaatgga cgaagcaggg attctgcaaa ccctatgcta ctccgtcaag ccgtcaattg
     4681 tctgattcgt taccaattat gacaacttga cggctacatc attcactttt tcttcacaac
     4741 cggcacggaa ctcgctcggg ctggccccgg tgcatttttt aaatacccgc gagaaataga
     4801 gttgatcgtc aaaaccaaca ttgcgaccga cggtggcgat aggcatccgg gtggtgctca
     4861 aaagcagctt cgcctggctg atacgttggt cctcgcgcca gcttaagacg ctaatcccta
     4921 actgctggcg gaaaagatgt gacagacgcg acggcgacaa gcaaacatgc tgtgcgacgc
     4981 tggcgatatc aaaattgctg tctgccaggt gatcgctgat gtactgacaa gcctcgcgta
     5041 cccgattatc catcggtgga tggagcgact cgttaatcgc ttccatgcgc cgcagtaaca
     5101 attgctcaag cagatttatc gccagcagct ccgaatagcg cccttcccct tgcccggcgt
     5161 taatgatttg cccaaacagg tcgctgaaat gcggctggtg cgcttcatcc gggcgaaaga
     5221 accccgtatt ggcaaatatt gacggccagt taagccattc atgccagtag gcgcgcggac
     5281 gaaagtaaac ccactggtga taccattcgc gagcctccgg atgacgaccg tagtgatgaa
     5341 tctctcctgg cgggaacagc aaaatatcac ccggtcggca aacaaattct cgtccctgat
     5401 ttttcaccac cccctgaccg cgaatggtga gattgagaat ataacctttc attcccagcg
     5461 gtcggtcgat aaaaaaatcg agataaccgt tggcctcaat cggcgttaaa cccgccacca
     5521 gatgggcatt aaacgagtat cccggcagca ggggatcatt ttgcgcttca gccatacttt
     5581 tcatactccc gccattcaga g
//
