LOCUS       sequence-151251-        4773 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 4773)
  AUTHORS   Alford SC, Ding Y, Simmen T, Campbell RE
  TITLE     Dimerization-dependent green and yellow fluorescent proteins.
  JOURNAL   ACS Synth Biol. 2012 Dec 21;1(12):569-75. doi: 10.1021/sb300050j. 
            Epub 2012 Aug 14.
  PUBMED    23656278
REFERENCE   2  (bases 1 to 4773)
  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"; doi: "10.1021/sb300050j";
            journalName: "ACS Synth Biol"; date: "2012-12-21- 21"; volume: "1"; 
            issue: "12"; pages: "569-75"
COMMENT     Sequence Label: pDDGFP-A
FEATURES             Location/Qualifiers
     source          1..4773
                     /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(560..578)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     828..847
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     1022..1045
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1174..1191)
                     /label=pBAD Reverse
                     /note="For vectors with E. coli araBAD promoter, reverse 
                     primer"
     terminator      1344..1430
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1522..1549
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1568..1659
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1660..1728,1729..2520)
                     /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(1878..1897)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2691..3279
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3180..3199
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3433..3450
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3465..3605
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3531..3550)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3691..3713
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(3869..4747)
                     /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 tcgagaatgg cgagcaagag cgaggaggtc atcaaagagt tcatgcgctt caaggtgcgc
      481 ttggagggct ccatgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgccct
      541 tacgagggca cccagaccgc caagctgaag gtgaccaagg gcggccccct gcccttcgcc
      601 tgggacatcc tgtcccccct gatcatgtac ggctccaaga tgtacgtgaa gcaccccgcc
      661 gacgtccccg attacatgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg
      721 cacttcgagg acggcggtct ggtgaccgcg acacaggact cctccctgca ggacggcacg
      781 ctgatctaca aggtgaagat gcgcggcacc aacttccccc ccgacggccc cgtaatgcag
      841 aagaagacct tgggctggga ttatgccacc gagcgcctgt accccgaaga aggcgtgctg
      901 aagggcgagc tcctggggcg cctgaagctg aaggacggcg gcctcaacct ggtggagtcc
      961 aagaccatct acatggccaa gaagcccgtg caactgcccg gctactactt cgtggacacc
     1021 aagctggaca tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgctcc
     1081 gagggccgcc accacctggg gcatggtaca ggtagcacag gcagcggtag ctccggtacc
     1141 taactgtttt ggcggatgag agaagatttt cagcctgata cagattaaat cagaacgcag
     1201 aagcggtctg ataaaacaga atttgcctgg cggcagtagc gcggtggtcc cacctgaccc
     1261 catgccgaac tcagaagtga aacgccgtag cgccgatggt agtgtggggt ctccccatgc
     1321 gagagtaggg aactgccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct
     1381 ttcgttttat ctgttgtttg tcggtgaacg ctctcctgag taggacaaat ccgccgggag
     1441 cggatttgaa cgttgcgaag caacggcccg gagggtggcg ggcaggacgc ccgccataaa
     1501 ctgccaggca tcaaattaag cagaaggcca tcctgacgga tggccttttt gcgtttctac
     1561 aaactctttt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     1621 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     1681 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     1741 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     1801 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     1861 agcactttta aagttctgct atgtggcgcg gtattatccc gtgttgacgc cgggcaagag
     1921 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     1981 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     2041 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     2101 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     2161 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     2221 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     2281 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     2341 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     2401 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     2461 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     2521 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     2581 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     2641 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     2701 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     2761 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     2821 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     2881 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     2941 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3001 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3061 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3121 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3181 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3241 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3301 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     3361 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     3421 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     3481 ctccttacgc atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc
     3541 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     3601 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3661 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     3721 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg
     3781 cataatgtgc ctgtcaaatg gacgaagcag ggattctgca aaccctatgc tactccgtca
     3841 agccgtcaat tgtctgattc gttaccaatt atgacaactt gacggctaca tcattcactt
     3901 tttcttcaca accggcacgg aactcgctcg ggctggcccc ggtgcatttt ttaaataccc
     3961 gcgagaaata gagttgatcg tcaaaaccaa cattgcgacc gacggtggcg ataggcatcc
     4021 gggtggtgct caaaagcagc ttcgcctggc tgatacgttg gtcctcgcgc cagcttaaga
     4081 cgctaatccc taactgctgg cggaaaagat gtgacagacg cgacggcgac aagcaaacat
     4141 gctgtgcgac gctggcgata tcaaaattgc tgtctgccag gtgatcgctg atgtactgac
     4201 aagcctcgcg tacccgatta tccatcggtg gatggagcga ctcgttaatc gcttccatgc
     4261 gccgcagtaa caattgctca agcagattta tcgccagcag ctccgaatag cgcccttccc
     4321 cttgcccggc gttaatgatt tgcccaaaca ggtcgctgaa atgcggctgg tgcgcttcat
     4381 ccgggcgaaa gaaccccgta ttggcaaata ttgacggcca gttaagccat tcatgccagt
     4441 aggcgcgcgg acgaaagtaa acccactggt gataccattc gcgagcctcc ggatgacgac
     4501 cgtagtgatg aatctctcct ggcgggaaca gcaaaatatc acccggtcgg caaacaaatt
     4561 ctcgtccctg atttttcacc accccctgac cgcgaatggt gagattgaga atataacctt
     4621 tcattcccag cggtcggtcg ataaaaaaat cgagataacc gttggcctca atcggcgtta
     4681 aacccgccac cagatgggca ttaaacgagt atcccggcag caggggatca ttttgcgctt
     4741 cagccatact tttcatactc ccgccattca gag
//
