LOCUS       Exported                5444 bp ds-DNA     circular SYN 04-JAN-2022
DEFINITION  Localization: Protein Expression Vector , Excitation: 604, Emission:
            659.
ACCESSION   .
VERSION     .
KEYWORDS    mCardinal-pBAD
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5444)
  AUTHORS   Chu J, Haynes RD, Corbel SY, Li P, Gonzalez-Gonzalez E, Burg JS, 
            Ataie NJ, Lam AJ, Cranfill PJ, Baird MA, Davidson MW, Ng HL, Garcia 
            KC, Contag CH, Shen K, Blau HM, Lin MZ
  TITLE     Non-invasive intravital imaging of cellular differentiation with a 
            bright red-excitable fluorescent protein.
  JOURNAL   Nat Methods. 2014 Mar 16. doi: 10.1038/nmeth.2888.
  PUBMED    24633408
REFERENCE   2  (bases 1 to 5444)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 4, 2022 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5444
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     RBS             1..23
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             43..60
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             64..96
                     /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             109..129
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             136..870
                     /codon_start=1
                     /product="far-red fluorescent protein derived from 
                     mNeptune, with further red-shifted spectra"
                     /label=mCardinal
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTTEGEGKPYEGTQTQ
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEATTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     primer_bind     complement(916..933)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1086..1172
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1264..1291
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1310..1401
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1402..2262
                     /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(1620..1639)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2304..2759
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2391..2410)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2601..2622
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     rep_origin      2870..3458
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3359..3378
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3612..3629
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3644..3784
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3710..3729)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3870..3892
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4030..4049
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
     CDS             complement(4229..5107)
                     /codon_start=1
                     /gene="araC"
                     /product="L-arabinose regulatory protein"
                     /label=araC
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     promoter        5134..5418
                     /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     5341..5360
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
ORIGIN
        1 tttgtttaac tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat
       61 ggtatggcta gcatgactgg tggacagcaa atgggtcggg atctgtacga gaacctgtac
      121 ttccagggct cgagcatggt gagcaagggc gaggagctga tcaaggagaa catgcacatg
      181 aagctgtaca tggaaggcac cgtgaacaac caccacttca agtgcaccac cgaaggggag
      241 ggcaagccct acgagggcac ccagacccag aggattaagg tggtggaggg aggccccctg
      301 ccgttcgcat tcgacatcct ggccacctgc tttatgtacg ggagcaagac cttcatcaac
      361 cacacccagg gcatccccga tttctttaag cagtccttcc ctgagggctt cacatgggag
      421 agagtcacca catacgaaga cgggggcgtg cttaccgtta cccaggacac cagcctccag
      481 gacggctgct tgatctacaa cgtcaagctc agaggggtga acttcccatc caacggccct
      541 gtgatgcaga agaaaacact cggctgggag gccaccaccg agaccctgta ccccgctgac
      601 ggcggcctgg aaggcagatg cgacatggcc ctgaagctcg tgggcggggg ccacctgcac
      661 tgcaacctga agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc
      721 gtctactttg tggaccgcag actggaaaga atcaaggagg ccgacaatga gacctacgtc
      781 gagcagcacg aggtggctgt ggccagatac tgcgacctcc ctagcaaact ggggcacaaa
      841 cttaatggca tggacgagct gtacaagtaa gaattcgaag cttggctgtt ttggcggatg
      901 agagaagatt ttcagcctga tacagattaa atcagaacgc agaagcggtc tgataaaaca
      961 gaatttgcct ggcggcagta gcgcggtggt cccacctgac cccatgccga actcagaagt
     1021 gaaacgccgt agcgccgatg gtagtgtggg gtctccccat gcgagagtag ggaactgcca
     1081 ggcatcaaat aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt
     1141 tgtcggtgaa cgctctcctg agtaggacaa atccgccggg agcggatttg aacgttgcga
     1201 agcaacggcc cggagggtgg cgggcaggac gcccgccata aactgccagg catcaaatta
     1261 agcagaaggc catcctgacg gatggccttt ttgcgtttct acaaactctt ttgtttattt
     1321 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     1381 taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct tattcccttt
     1441 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     1501 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     1561 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     1621 ctatgtggcg cggtattatc ccgtgttgac gccgggcaag agcaactcgg tcgccgcata
     1681 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     1741 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     1801 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     1861 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     1921 gacgagcgtg acaccacgat gcctgcagca atggcaacaa cgttgcgcaa actattaact
     1981 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     2041 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatct
     2101 ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga tggtaagccc
     2161 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     2221 cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga ccaagtttac
     2281 tcatatatac tttagattga tttacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2341 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2401 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2461 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgattt
     2521 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2581 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     2641 ctcgggctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     2701 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgt ttacaattta
     2761 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     2821 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     2881 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     2941 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3001 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3061 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3121 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3181 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     3241 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     3301 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     3361 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     3421 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     3481 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     3541 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     3601 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcctgatg cggtattttc
     3661 tccttacgca tctgtgcggt atttcacacc gcatctggtg cactctcagt acaatctgct
     3721 ctgatgccgc atagttaagc cagtatacac tccgctatcg ctacgtgact gggtcatggc
     3781 tgcgccccga cacccgccaa cacccgctga cgcgccctga cgggcttgtc tgctcccggc
     3841 atccgcttac agacaagctg tgaccgtctc cgggagctgc atgtgtcaga ggttttcacc
     3901 gtcatcaccg aaacgcgcga ggcagaagga gatggcgccc aacagtcccc cggccacggg
     3961 gcctgccacc atacccacgc cgaaacaagc gctcatgagc ccgaagtggc gagcccgatc
     4021 ttccccatcg gtgatgtcgg cgatataggc gccagcaacc gcacctgtgg cgccggtgat
     4081 gccggccacg atgcgtccgg cgtagaggat ctgctcatgt ttgacagctt atcatcgatg
     4141 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
     4201 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
     4261 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     4321 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     4381 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     4441 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     4501 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     4561 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     4621 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     4681 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     4741 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     4801 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     4861 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     4921 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     4981 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     5041 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     5101 cagccatact tttcatactc ccgccattca gagaagaaac caattgtcca tattgcatca
     5161 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc
     5221 ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa
     5281 gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct
     5341 atgccatagc atttttatcc ataagattag cggatactac ctgacgcttt ttatcgcaac
     5401 tctctactgt ttctccatac ccgttttttt gggctagaaa taat
//
