LOCUS       62056_22015        4859 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4859)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    106..127
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        142..172
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    180..196
                     /label=lac operator
                     /note="The lac repressor binds to the lac operator to
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     204..220
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             293..1024
                     /codon_start=1
                     /label=mNeptune2
                     /note="far-red fluorescent protein with spectra similar to
                     those of mNeptune but improved brightness"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTG
                     RIKVVEGGPLPFAFDILATCFMYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTVTQDTSLQDGCLIYNVKLRGVNFPSNGPVMQKKTLGWEASTETLYPADGGLEGRCD
                     MALKLVGGGHLHCNLKTTYRSKKPAKNLKMPGVYFVDRRLERIKEADNETYVEQHEVAV
                     ARYCDLPSKLGHKLNGMDELYK"
     protein_bind    complement(1025..1058)
                     /label=lox66
                     /note="Right element (RE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1059..1283
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        1284..1783
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             1793..2188
                     /codon_start=1
                     /label=BSD
                     /note="blasticidin S deaminase"
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG"
     protein_bind    complement(2417..2450)
                     /label=lox71
                     /note="Left element (LE) mutant of loxP (Araki et al.,
                     2010). Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     complement(2463..2479)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        2953..3057
                     /label=AmpR promoter
     CDS             3058..3915
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      4089..4677
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
      121 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
      181 tgtgagcgga taacaatttc acacaggaaa cagctatgac catgattacg ccaagcttga
      241 gggcagagga agtcttctaa catgcggtga cgtggaggag aatcccggcc ctatggtgtc
      301 taagggcgaa gagctgatta aggagaacat gcacatgaag ctgtacatgg agggcaccgt
      361 gaacaaccac cacttcaagt gcacatccga gggcgaaggc aagccctacg agggcaccca
      421 gaccggcaga atcaaggtgg tcgagggcgg ccctctcccc ttcgccttcg acatcctggc
      481 tacctgcttc atgtacggca gcaagacctt catcaaccac acccagggca tccccgattt
      541 ctttaagcag tccttccctg agggcttcac atgggagaga gtcaccacat acgaagacgg
      601 gggcgtgctt accgttaccc aggacaccag cctccaggac ggctgcttga tctacaacgt
      661 caagctcaga ggggtgaact tcccatccaa cggccctgtg atgcagaaga aaacactcgg
      721 ctgggaggcc agtaccgaga ccctgtaccc cgctgacggc ggcctggaag gcagatgcga
      781 catggccctg aagctcgtgg gcgggggcca cctgcactgc aacctgaaga ccacatacag
      841 atccaagaaa cccgctaaga acctcaagat gcccggcgtc tactttgtgg accgcagact
      901 ggaaagaatc aaggaggccg acaatgagac ctacgtcgag cagcacgagg tggctgtggc
      961 cagatactgc gacctcccta gcaaactggg gcacaaactt aatggcatgg acgagctgta
     1021 caagataact tcgtatagca tacattatac gaacggtact gtgccttcta gttgccagcc
     1081 atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt
     1141 cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct
     1201 ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc
     1261 tggggatgcg gtgggctcta tgggggtagg ggaggcgctt ttcccaaggc agtctggagc
     1321 atgcgcttta gcagccccgc tgggcacttg gcgctacaca agtggcctct ggcctcgcac
     1381 acattccaca tccaccggta ggcgccaacc ggctccgttc tttggtggcc ccttcgcgcc
     1441 accttctact cctcccctag tcaggaagtt cccccccgcc ccgcagctcg cgtcgtgcag
     1501 gacgtgacaa atggaagtag cacgtctcac tagtctcgtg cagatggaca gcaccgctga
     1561 gcaatggaag cgggtaggcc tttggggcag cggccaatag cagctttgct ccttcgcttt
     1621 ctgggctcag aggctgggaa ggggtgggtc cgggggcggg ctcaggggcg ggctcagggg
     1681 cggggcgggc gcccgaaggt cctccggagg cccggcattc tgcacgcttc aaaagcgcac
     1741 gtctgccgcg ctgttctcct cttcctcatc tccgggcctt tcgaggctcg agatggccaa
     1801 gcctttgtct caagaagaat ccaccctcat tgaaagagca acggctacaa tcaacagcat
     1861 ccccatctct gaagactaca gcgtcgccag cgcagctctc tctagcgacg gccgcatctt
     1921 cactggtgtc aatgtatatc attttactgg gggaccttgt gcagaactcg tggtgctggg
     1981 cactgctgct gctgcggcag ctggcaacct gacttgtatc gtcgcgatcg gaaatgagaa
     2041 caggggcatc ttgagcccct gcggacggtg ccgacaggtg cttctcgatc tgcatcctgg
     2101 gatcaaagcc atagtgaagg acagtgatgg acagccgacg gcagttggga ttcgtgaatt
     2161 gctgccctct ggttatgtgt gggagggcta actgtgcctt ctagttgcca gccatctgtt
     2221 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2281 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2341 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2401 gcggtgggct ctatggtacc gttcgtatag catacattat acgaagttat gagctcgaat
     2461 tcactggccg tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat
     2521 cgccttgcag cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat
     2581 cgcccttccc aacagttgcg cagcctgaat ggcgaatggc gcctgatgcg gtattttctc
     2641 cttacgcatc tgtgcggtat ttcacaccgc atatggtgca ctctcagtac aatctgctct
     2701 gatgccgcat agttaagcca gccccgacac ccgccaacac ccgctgacgc gccctgacgg
     2761 gcttgtctgc tcccggcatc cgcttacaga caagctgtga ccgtctccgg gagctgcatg
     2821 tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc
     2881 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2941 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     3001 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     3061 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     3121 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     3181 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     3241 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     3301 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     3361 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     3421 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     3481 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     3541 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     3601 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3661 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3721 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3781 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3841 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3901 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3961 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     4021 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4081 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4141 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4201 actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc
     4261 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4321 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4381 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4441 cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt
     4501 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4561 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4621 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4681 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4741 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4801 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaaga
//
