LOCUS       sequence-63488-e        5010 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 5010)
  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 5010)
  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-iRFP670
FEATURES             Location/Qualifiers
     source          1..5010
                     /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..1365
                     /codon_start=1
                     /product="near-infrared fluorescent protein with an 
                     emission peak at 670 nm, engineered from a bacterial 
                     phytochrome (Shcherbakova and Verkhusha, 2013)"
                     /label=iRFP670
                     /note="derived from Rhodopseudomonas palustris RpBphP6"
                     /translation="MPRKVDLTSCDREPIHIPGSIQPCGCLLACDAQAVRITRITENAG
                     AFFGRETPRVGELLADYFGETEAHALRNALAQSSDPKRPALIFGWRDGLTGRTFDISLH
                     RHDGTSIIEFEPAAAEQADNPLRLTRQIIARTKELKSLEEMAARVPRYLQAMLGYHRVM
                     LYRFADDGSGMVIGEAKRSDLESFLGQHFPASLVPQQARLLYLKNAIRVVSDSRGISSR
                     IVPEHDASGAALDLSFAHLRSISPCHLEFLRNMGVSASMSLSIIIDGTLWGLIICHHYE
                     PRAVPMAQRVAAEMFADFLSLHFTAAHHQR"
     primer_bind     complement(1411..1428)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1581..1667
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1759..1786
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1805..1896
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1897..1965,1966..2757)
                     /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(2115..2134)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2928..3516
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3417..3436
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3670..3687
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3702..3842
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3768..3787)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3928..3950
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4106..4984)
                     /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 tgccgcgtaa ggtcgatctc acctcctgcg atcgcgagcc gatccacatc
      481 cccggcagca ttcagccgtg cggctgcctg ctagcctgcg acgcgcaggc ggtgcggatc
      541 acgcgcatta cggaaaatgc cggcgcgttc tttggacgcg aaactccgcg ggtcggtgag
      601 ctactcgccg attacttcgg cgagaccgaa gcccatgcgc tgcgcaacgc actggcgcag
      661 tcctccgatc caaagcgacc ggcgctgatc ttcggttggc gcgacggcct gaccggccgc
      721 accttcgaca tctcactgca tcgccatgac ggtacatcga tcatcgagtt cgagcctgcg
      781 gcggccgaac aggccgacaa tccgctgcgg ctgacgcggc agatcatcgc gcgcaccaaa
      841 gaactgaagt cgctcgaaga gatggccgca cgggtgccgc gctatctgca ggcgatgctc
      901 ggctatcacc gcgtgatgtt gtaccgcttc gcggacgacg gctccgggat ggtgatcggc
      961 gaggcgaagc gcagcgacct cgagagcttt ctcggtcagc actttccggc gtcgctggtc
     1021 ccgcagcagg cgcggctact gtacttgaag aacgcgatcc gcgtggtctc ggattcgcgc
     1081 ggcatcagca gccggatcgt gcccgagcac gacgcctccg gcgccgcgct cgatctgtcg
     1141 ttcgcgcacc tgcgcagcat ctcgccctgc catctcgaat ttctgcggaa catgggcgtc
     1201 agcgcctcga tgtcgctgtc gatcatcatt gacggcacgc tatggggatt gatcatctgt
     1261 catcattacg agccgcgtgc cgtgccgatg gcgcagcgcg tcgcggccga aatgttcgcc
     1321 gacttcttat cgctgcactt caccgccgcc caccaccaac gctaagaatt cgaagcttgg
     1381 ctgttttggc ggatgagaga agattttcag cctgatacag attaaatcag aacgcagaag
     1441 cggtctgata aaacagaatt tgcctggcgg cagtagcgcg gtggtcccac ctgaccccat
     1501 gccgaactca gaagtgaaac gccgtagcgc cgatggtagt gtggggtctc cccatgcgag
     1561 agtagggaac tgccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc
     1621 gttttatctg ttgtttgtcg gtgaacgctc tcctgagtag gacaaatccg ccgggagcgg
     1681 atttgaacgt tgcgaagcaa cggcccggag ggtggcgggc aggacgcccg ccataaactg
     1741 ccaggcatca aattaagcag aaggccatcc tgacggatgg cctttttgcg tttctacaaa
     1801 ctcttttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     1861 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     1921 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     1981 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     2041 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     2101 acttttaaag ttctgctatg tggcgcggta ttatcccgtg ttgacgccgg gcaagagcaa
     2161 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     2221 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     2281 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     2341 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     2401 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     2461 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     2521 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     2581 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     2641 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     2701 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     2761 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     2821 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     2881 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     2941 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     3001 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     3061 ataccaaata ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     3121 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     3181 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     3241 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     3301 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     3361 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     3421 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     3481 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     3541 cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat
     3601 tctgtggata accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg
     3661 accgagcgca gcgagtcagt gagcgaggaa gcggaagagc gcctgatgcg gtattttctc
     3721 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     3781 gatgccgcat agttaagcca gtatacactc cgctatcgct acgtgactgg gtcatggctg
     3841 cgccccgaca cccgccaaca cccgctgacg cgccctgacg ggcttgtctg ctcccggcat
     3901 ccgcttacag acaagctgtg accgtctccg ggagctgcat gtgtcagagg ttttcaccgt
     3961 catcaccgaa acgcgcgagg cagcagatca attcgcgcgc gaaggcgaag cggcatgcat
     4021 aatgtgcctg tcaaatggac gaagcaggga ttctgcaaac cctatgctac tccgtcaagc
     4081 cgtcaattgt ctgattcgtt accaattatg acaacttgac ggctacatca ttcacttttt
     4141 cttcacaacc ggcacggaac tcgctcgggc tggccccggt gcatttttta aatacccgcg
     4201 agaaatagag ttgatcgtca aaaccaacat tgcgaccgac ggtggcgata ggcatccggg
     4261 tggtgctcaa aagcagcttc gcctggctga tacgttggtc ctcgcgccag cttaagacgc
     4321 taatccctaa ctgctggcgg aaaagatgtg acagacgcga cggcgacaag caaacatgct
     4381 gtgcgacgct ggcgatatca aaattgctgt ctgccaggtg atcgctgatg tactgacaag
     4441 cctcgcgtac ccgattatcc atcggtggat ggagcgactc gttaatcgct tccatgcgcc
     4501 gcagtaacaa ttgctcaagc agatttatcg ccagcagctc cgaatagcgc ccttcccctt
     4561 gcccggcgtt aatgatttgc ccaaacaggt cgctgaaatg cggctggtgc gcttcatccg
     4621 ggcgaaagaa ccccgtattg gcaaatattg acggccagtt aagccattca tgccagtagg
     4681 cgcgcggacg aaagtaaacc cactggtgat accattcgcg agcctccgga tgacgaccgt
     4741 agtgatgaat ctctcctggc gggaacagca aaatatcacc cggtcggcaa acaaattctc
     4801 gtccctgatt tttcaccacc ccctgaccgc gaatggtgag attgagaata taacctttca
     4861 ttcccagcgg tcggtcgata aaaaaatcga gataaccgtt ggcctcaatc ggcgttaaac
     4921 ccgccaccag atgggcatta aacgagtatc ccggcagcag gggatcattt tgcgcttcag
     4981 ccatactttt catactcccg ccattcagag
//
