LOCUS       Exported                7831 bp ds-DNA     circular SYN 05-OCT-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7831)
  AUTHORS   Carissa Grose
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Oct 7, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7831
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        28..119
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     CDS             176..1273
                     /codon_start=1
                     /gene="malE (mutated)"
                     /product="maltose binding protein from E. coli"
                     /label=MBP
                     /note="This version of the gene does not encode a signal 
                     sequence, so MBP will remain in the cytosol."
                     /translation="KIEEGKLVIWINGDKGYNGLAEVGKKFEKDTGIKVTVEHPDKLEE
                     KFPQVAATGDGPDIIFWAHDRFGGYAQSGLLAEITPDKAFQDKLYPFTWDAVRYNGKLI
                     AYPIAVEALSLIYNKDLLPNPPKTWEEIPALDKELKAKGKSALMFNLQEPYFTWPLIAA
                     DGGYAFKYENGKYDIKDVGVDNAGAKAGLTFLVDLIKNKHMNADTDYSIAEAAFNKGET
                     AMTINGPWAWSNIDTSKVNYGVTVLPTFKGQPSKPFVGVLSAGINAASPNKELAKEFLE
                     NYLLTDEGLEAVNKDKPLGAVALKSYEEELAKDPRIAATMENAQKGEIMPNIPQMSAFW
                     YAVRTAVINAASGRQTVDEALKDAQT"
     regulatory      1130..1139
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     protein_bind    1292..1416
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        1441..1471
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             1525..2184
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             2526..2831
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VSRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     protein_bind    complement(2872..2996)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     polyA_signal    3141..3275
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  3304..3469
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      3653..4108
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4135..4239
                     /gene="bla"
                     /label=AmpR promoter
     CDS             4240..5100
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5271..5859
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  6162..6386
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(6453..6986)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(7175..7203)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     polyA_signal    7712..7760
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     complement(join(7811..7831,1..5))
                     /label=21546
     primer_bind     join(7828..7831,1..27)
                     /label=21545
ORIGIN
        1 tcgaggtata ctccggaata ttaatagatc atggagataa ttaaaatgat aaccatctcg
       61 caaataaata agtattttac tgttttcgta acagttttgt aataaaaaaa cctataaata
      121 ttccggatta ttcataccgt cccaccatcg ggcgcggatc accatgggat ccggcaaaat
      181 cgaagaaggt aaactggtaa tctggattaa cggcgataaa ggctataacg gtctcgctga
      241 agtcggtaag aaattcgaga aagataccgg aattaaagtc accgttgagc atccggataa
      301 actggaagag aaattcccac aggttgcggc aactggcgat ggccctgaca ttatcttctg
      361 ggcacacgac cgctttggtg gctacgctca atctggcctg ttggctgaaa tcaccccgga
      421 caaagcgttc caggacaagc tgtatccgtt tacctgggat gccgtacgtt acaacggcaa
      481 gctgattgct tacccgatcg ctgttgaagc gttatcgctg atttataaca aagatctgct
      541 gccgaacccg ccaaaaacct gggaagagat cccggcgctg gataaagaac tgaaagcgaa
      601 aggtaagagc gcgctgatgt tcaacctgca agaaccgtac ttcacctggc cgctgattgc
      661 tgctgacggg ggttatgcgt tcaagtatga aaacggcaag tacgacatta aagacgtggg
      721 cgtggataac gctggcgcga aagcgggtct gaccttcctg gttgacctga ttaaaaacaa
      781 acacatgaat gcagacaccg attactccat cgcagaagct gcctttaata aaggcgaaac
      841 agcgatgacc atcaacggcc cgtgggcatg gtccaacatc gacaccagca aagtgaatta
      901 tggtgtaacg gtactgccga ccttcaaggg tcaaccatcc aaaccgttcg ttggcgtgct
      961 gagcgcaggt attaacgccg ccagtccgaa caaagagctg gcaaaagagt tcctcgaaaa
     1021 ctatctgctg actgatgaag gtctggaagc ggttaataaa gacaaaccgc tgggtgccgt
     1081 agcgctgaag tcttacgagg aagagttggc gaaagatcca cgtattgccg ccaccatgga
     1141 aaacgcccag aaaggtgaaa tcatgccgaa catcccgcag atgtccgctt tctggtatgc
     1201 cgtgcgtact gcggtgatca acgccgccag cggtcgtcag actgtcgatg aagccctgaa
     1261 agacgcgcag actaattcgg gatcctcgat cacaagtttg tacaaaaaag ctgaacgaga
     1321 aacgtaaaat gatataaata tcaatatatt aaattagatt ttgcataaaa aacagactac
     1381 ataatactgt aaaacacaac atatccagtc actatggcgg ccgcattagg caccccaggc
     1441 tttacacttt atgcttccgg ctcgtataat gtgtggattt tgagttagga tccggcgaga
     1501 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
     1561 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
     1621 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
     1681 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
     1741 ttccgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
     1801 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
     1861 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
     1921 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     1981 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     2041 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     2101 catgccgtct gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     2161 gatgagtggc agggcggggc gtaaacgcgt ggatccggct tactaaaagc cagataacag
     2221 tatgcgtatt tgcgcgctga tttttgcggt ataagaatat atactgatat gtatacccga
     2281 agtatgtcaa aaagaggtgt gctatgaagc agcgtattac agtgacagtt gacagcgaca
     2341 gctatcagtt gctcaaggca tatatgatgt caatatctcc ggtctggtaa gcacaaccat
     2401 gcagaatgaa gcccgtcgtc tgcgtgccga acgctggaaa gcggaaaatc aggaagggat
     2461 ggctgaggtc gcccggttta ttgaaatgaa cggctctttt gctgacgaga acagggactg
     2521 gtgaaatgca gtttaaggtt tacacctata aaagagagag ccgttatcgt ctgtttgtgg
     2581 atgtacagag tgatattatt gacacgcccg ggcgacggat ggtgatcccc ctggccagtg
     2641 cacgtctgct gtcagataaa gtctcccgtg aactttaccc ggtggtgcat atcggggatg
     2701 aaagctggcg catgatgacc accgatatgg ccagtgtgcc ggtctccgtt atcggggaag
     2761 aagtggctga tctcagccac cgcgaaaatg acatcaaaaa cgccattaac ctgatgttct
     2821 ggggaatata aatgtcaggc tccgttatac acagccagtc tgcaggtcga ccatagtgac
     2881 tggatatgtt gtgttttaca gtattatgta gtctgttttt tatgcaaaat ctaatttaat
     2941 atattgatat ttatatcatt ttacgtttct cgttcagctt tcttgtacaa agtggtgatc
     3001 gagctagcta gaagcttgtc gagaagtact agaggatcat aatcagccat accacatttg
     3061 tagaggtttt acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa
     3121 tgaatgcaat tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca
     3181 atagcatcac aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt
     3241 ccaaactcat caatgtatct tatcatgtct ggatctgatc actgcttgag cctaggagat
     3301 ccgaaccaga taagtgaaat ctagttccaa actattttgt catttttaat tttcgtatta
     3361 gcttacgacg ctacacccag ttcccatcta ttttgtcact cttccctaaa taatccttaa
     3421 aaactccatt tccacccctc ccagttccca actattttgt ccgcccacag cggggcattt
     3481 ttcttcctgt tatgttttta atcaaacatc ctgccaactc catgtgacaa accgtcatct
     3541 tcggctactt tttctctgtc acagaatgaa aatttttctg tcatctcttc gttattaatg
     3601 tttgtaattg actgaatatc aacgcttatt tgcagcctga atggcgaatg ggacgcgccc
     3661 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     3721 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     3781 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     3841 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     3901 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     3961 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     4021 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     4081 tttaacaaaa tattaacgtt tacaatttca ggtggcactt ttcggggaaa tgtgcgcgga
     4141 acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa
     4201 ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca acatttccgt
     4261 gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca cccagaaacg
     4321 ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta catcgaactg
     4381 gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt tccaatgatg
     4441 agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc cgggcaagag
     4501 caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc accagtcaca
     4561 gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc cataaccatg
     4621 agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa ggagctaacc
     4681 gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga accggagctg
     4741 aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat ggcaacaacg
     4801 ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca attaatagac
     4861 tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc ggctggctgg
     4921 tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat tgcagcactg
     4981 gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag tcaggcaact
     5041 atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa gcattggtaa
     5101 ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca tttttaattt
     5161 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     5221 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     5281 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     5341 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     5401 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     5461 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     5521 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     5581 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     5641 ctgagatacc tacagcgtga gcattgagaa agcgccacgc ttcccgaagg gagaaaggcg
     5701 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     5761 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5821 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5881 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5941 gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg ccgcagccga
     6001 acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcctgat gcggtatttt
     6061 ctccttacgc atctgtgcgg tatttcacac cgcagaccag ccgcgtaacc tggcaaaatc
     6121 ggttacggtt gagtaataaa tggatgccct gcgtaagcgg gtgtgggcgg acaataaagt
     6181 cttaaactga acaaaataga tctaaactat gacaataaag tcttaaacta gacagaatag
     6241 ttgtaaactg aaatcagtcc agttatgctg tgaaaaagca tactggactt ttgttatggc
     6301 taaagcaaac tcttcatttt ctgaagtgca aattgcccgt cgtattaaag aggggcgtgg
     6361 ccaagggcat ggtaaagact atattcgcgg cgttgtgaca atttaccgaa caactccgcg
     6421 gccgggaagc cgatctcggc ttgaacgaat tgttaggtgg cggtacttgg gtcgatatca
     6481 aagtgcatca cttcttcccg tatgcccaac tttgtataga gagccactgc gggatcgtca
     6541 ccgtaatctg cttgcacgta gatcacataa gcaccaagcg cgttggcctc atgcttgagg
     6601 agattgatga gcgcggtggc aatgccctgc ctccggtgct cgccggagac tgcgagatca
     6661 tagatataga tctcactacg cggctgctca aacctgggca gaacgtaagc cgcgagagcg
     6721 ccaacaaccg cttcttggtc gaaggcagca agcgcgatga atgtcttact acggagcaag
     6781 ttcccgaggt aatcggagtc cggctgatgt tgggagtagg tggctacgtc tccgaactca
     6841 cgaccgaaaa gatcaagagc agcccgcatg gatttgactt ggtcagggcc gagcctacat
     6901 gtgcgaatga tgcccatact tgagccacct aactttgttt tagggcgact gccctgctgc
     6961 gtaacatcgt tgctgctgcg taacatcgtt gctgctccat aacatcaaac atcgacccac
     7021 ggcgtaacgc gcttgctgct tggatgcccg aggcatagac tgtacaaaaa aacagtcata
     7081 acaagccatg aaaaccgcca ctgcgccgtt accaccgctg cgttcggtca aggttctgga
     7141 ccagttgcgt gagcgcatac gctacttgca ttacagttta cgaaccgaac aggcttatgt
     7201 caactgggtt cgtgccttca tccgtttcca cggtgtgcgt cacccggcaa ccttgggcag
     7261 cagcgaagtc gaggcatttc tgtcctggct ggcgaacgag cgcaaggttt cggtctccac
     7321 gcatcgtcag gcattggcgg ccttgctgtt cttctacggc aaggtgctgt gcacggatct
     7381 gccctggctt caggagatcg gtagacctcg gccgtcgcgg cgcttgccgg tggtgctgac
     7441 cccggatgaa gtggttcgca tcctcggttt tctggaaggc gagcatcgtt tgttcgccca
     7501 ggactctagc tatagttcta gtggttggcc tacgtacccg tagtggctat ggcagggctt
     7561 gccgccccga cgttggctgc gagccctggg ccttcacccg aacttggggg ttggggtggg
     7621 gaaaaggaag aaacgcgggc gtattggtcc caatggggtc tcggtggggt atcgacagag
     7681 tgccagccct gggaccgaac cccgcgttta tgaacaaacg acccaacacc cgtgcgtttt
     7741 attctgtctt tttattgccg tcatagcgcg ggttccttcc ggtattgtct ccttccgtgt
     7801 ttcagttagc ctcccccatc tcccggtacc c
//
