LOCUS       sequence-63489-e        5025 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 5025)
  AUTHORS   Shcherbakova DM, Verkhusha VV
  TITLE     Near-infrared fluorescent proteins for multicolor in vivo imaging.
  JOURNAL   Nat Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521.
  PUBMED    23770755
REFERENCE   2  (bases 1 to 5025)
  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 
            Methods. 2013 Jun 16. doi: 10.1038/nmeth.2521."
COMMENT     Sequence Label: pBAD/HisB-iRFP682
FEATURES             Location/Qualifiers
     source          1..5025
                     /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"
     CDS             430..1380
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 682 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP682
                     /note="derived from Rhodopseudomonas palustris RpBphP2"
                     /translation="MAEGSVARQPDLLTCDDEPIHIPGAIQPHGLLLALAADMTIVAGS
                     DNLPELTGLAIGALIGRSAADVFDSETHNRLTIALAEPGAAVGAPITVGFTMRKDAGFI
                     GSWHRHDQLIFLELEPPQRDVAEPQAFFRRTNSAIRRLQAAETLESACAAAAQEVRKIT
                     GFDRVMIYRFASDFSGVVIAEDRCAEVESKLGLHYPASAVPAQARRLYTINPVRIIPDI
                     NYRPVPVTPDLNPVTGRPIDLSFAILRSVSPCHLEFMRNIGMHGTMSISILRGERLWGL
                     IVCHHRTPYYVDLDGRQACELVAQVLAWQIGVMEE"
     primer_bind     complement(1426..1443)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1596..1682
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1774..1801
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1820..1911
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1912..1980,1981..2772)
                     /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(2130..2149)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2943..3531
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3432..3451
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3685..3702
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3717..3857
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3783..3802)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3943..3965
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4121..4999)
                     /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 tggcggaagg atccgtcgcc aggcagcctg acctcttgac ctgcgacgat
      481 gagccgatcc atatccccgg tgccatccaa ccgcatggac tgctgctcgc cctcgccgcc
      541 gacatgacga tcgttgccgg cagcgacaac cttcccgaac tcaccggact ggcgatcggc
      601 gccctgatcg gccgctctgc ggccgatgtc ttcgactcgg agacgcacaa ccgtctgacg
      661 atcgccttgg ccgagcccgg ggcggccgtc ggagcaccga tcactgtcgg cttcacgatg
      721 cgaaaggacg caggcttcat cggctcctgg catcgccatg atcagctcat cttcctcgag
      781 ctcgagcctc cccagcggga cgtcgccgag ccgcaggcgt tcttccgccg caccaacagc
      841 gccatccgcc gcctgcaggc cgccgaaacc ttggaaagcg cctgcgccgc cgcggcgcaa
      901 gaggtgcgga agattaccgg cttcgatcgg gtgatgatct atcgcttcgc ctccgacttc
      961 agcggggtgg tgatcgcaga ggatcgatgc gccgaggtcg agtcaaaact aggcctgcac
     1021 tatcctgcct cagcggtgcc ggcgcaggcc cgtcggctct ataccatcaa cccggtacgg
     1081 atcattcccg atatcaatta tcggccggtg ccggtcaccc cagacctcaa tccggtcacc
     1141 gggcggccga ttgatcttag cttcgccatc ctgcgcagcg tctcgccctg ccatttggag
     1201 ttcatgcgca acataggcat gcacggcacg atgtcgatct cgattttgcg cggcgagcga
     1261 ctgtggggat tgatcgtttg ccatcaccga acgccgtact acgtcgatct cgatggccgc
     1321 caagcctgcg agctagtcgc ccaggttctg gcctggcaga tcggcgtgat ggaagagtga
     1381 gaattcgaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1441 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1501 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1561 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1621 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1681 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1741 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1801 ttgcgtttct acaaactctt ttgtttattt ttctaaatac attcaaatat gtatccgctc
     1861 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt
     1921 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     1981 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     2041 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     2101 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtgttgac
     2161 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     2221 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     2281 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     2341 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     2401 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca
     2461 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     2521 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     2581 ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc
     2641 attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     2701 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     2761 aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt
     2821 catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc
     2881 ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct
     2941 tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta
     3001 ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc
     3061 ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac
     3121 ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct
     3181 gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat
     3241 aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg
     3301 acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa
     3361 gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg
     3421 gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga
     3481 cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc
     3541 aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct
     3601 gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct
     3661 cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgcctg
     3721 atgcggtatt ttctccttac gcatctgtgc ggtatttcac accgcatatg gtgcactctc
     3781 agtacaatct gctctgatgc cgcatagtta agccagtata cactccgcta tcgctacgtg
     3841 actgggtcat ggctgcgccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt
     3901 gtctgctccc ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc
     3961 agaggttttc accgtcatca ccgaaacgcg cgaggcagca gatcaattcg cgcgcgaagg
     4021 cgaagcggca tgcataatgt gcctgtcaaa tggacgaagc agggattctg caaaccctat
     4081 gctactccgt caagccgtca attgtctgat tcgttaccaa ttatgacaac ttgacggcta
     4141 catcattcac tttttcttca caaccggcac ggaactcgct cgggctggcc ccggtgcatt
     4201 ttttaaatac ccgcgagaaa tagagttgat cgtcaaaacc aacattgcga ccgacggtgg
     4261 cgataggcat ccgggtggtg ctcaaaagca gcttcgcctg gctgatacgt tggtcctcgc
     4321 gccagcttaa gacgctaatc cctaactgct ggcggaaaag atgtgacaga cgcgacggcg
     4381 acaagcaaac atgctgtgcg acgctggcga tatcaaaatt gctgtctgcc aggtgatcgc
     4441 tgatgtactg acaagcctcg cgtacccgat tatccatcgg tggatggagc gactcgttaa
     4501 tcgcttccat gcgccgcagt aacaattgct caagcagatt tatcgccagc agctccgaat
     4561 agcgcccttc cccttgcccg gcgttaatga tttgcccaaa caggtcgctg aaatgcggct
     4621 ggtgcgcttc atccgggcga aagaaccccg tattggcaaa tattgacggc cagttaagcc
     4681 attcatgcca gtaggcgcgc ggacgaaagt aaacccactg gtgataccat tcgcgagcct
     4741 ccggatgacg accgtagtga tgaatctctc ctggcgggaa cagcaaaata tcacccggtc
     4801 ggcaaacaaa ttctcgtccc tgatttttca ccaccccctg accgcgaatg gtgagattga
     4861 gaatataacc tttcattccc agcggtcggt cgataaaaaa atcgagataa ccgttggcct
     4921 caatcggcgt taaacccgcc accagatggg cattaaacga gtatcccggc agcaggggat
     4981 cattttgcgc ttcagccata cttttcatac tcccgccatt cagag
//
