LOCUS       Exported                6100 bp ds-DNA     circular SYN 05-OCT-2020
DEFINITION  R4-R3 baculovirus vector, no promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6100)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Oct 7, 2020 from SnapGene Viewer 5.0.8
            https://www.snapgene.com
COMMENT     Vector molecule:  pDest-600*
               Fragment ends: NruI
               Fragment size: 4393
            Insert molecule:  pSpcRFA43cassette
               Fragment ends: EcoRV
               Fragment size: 1707
            pDP600 was cut with Nru1.  This digest left a fair amount of 
            supercoiled vector. However, due to the fact that the insert has 
            additional antibiotic resistance in it, this was deemed okay.  
            Ligation to the 43 cassette yielded a lot of randomly sized clones, 
            and it was therefore decided that the insert was bad.  7/28/2008  JM
            A second tube of the RFA43 cassette was used, yielding the same 
            result, with small background colonies but none with the correct 
            insert.  7/31/2008 JM
            The glycerol stock was found for the RFA43 insert, and a maxi prep 
            was done on the parent vector.  However, upon running the new maxi 
            prep on a gel, it was discovered that it was  running at 7.5kB 
            instead of 4.4kB.  Therefore, this vector was put on hold until Dom 
            finds another stock for the RFA43 insert.  8/4/2008  JM
            An original tube of uncut pSpcFRA43 DNA was found, and the DNA was 
            Xfed into DH5alhpas, 2 colonies were picked and prepped.  They were 
            the correct 4.4 size, and a glycerol stock was made to replace the 
            old one.  8/14/2008 JM
            A maxi prep was done using this new glycerol stock, and a gel check 
            proved that this was still the correct 4.4 band.  An EcoR5 digest 
            was done on this new vector, and it was cleaned, thus giving a new 
            batch of insert.  8/18/2008 JM
            This new insert was used, in a 1:3 ratio with the Nru! cut pDP600 
            vector to do a ligation.  The ligation was Xfed into DB3.1, plated 
            on Amp/Chlor. 8/22/2008  JM
            5 colonies were picked from this plate, mini prepped, and run on a 
            gel.  They were extremely faint, however, they all appeared to be 
            the corrrect size.  They were digested with BamH1 to check 
            directionality, and the digests were also faint, but clones 1 and 3 
            appear correct, and were sent to sequencing.  8/25/2008  JM
            Clones 1 and 3 were both sequence verified on 9/10/08, wells E4-5 
            and F4-5.  Sequence of attR4 and attR3 and flanking regions are 
            correct.  DE 9/12/08
            Clone 1 was kept, and maxi prepped.  JM 9/28/2008
            The glycerol stock was used to start a 2mL culture in both A/C and 
            G/C to test for Gent resistance in the vector.  Both cultures grew 
            well, and the mini prep of both showed a clean band of the correct 
            size.  Therefore, this vector is Gent. resistant.  JM 9/29/2008
FEATURES             Location/Qualifiers
     source          1..6100
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     polyA_signal    129..263
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     mobile_element  292..457
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     rep_origin      641..1096
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1123..1227
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1228..2088
                     /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      2259..2847
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     mobile_element  3150..3374
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7R
                     /note="mini-Tn7 element (right end of the Tn7 transposon)"
     CDS             complement(3441..3974)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPRFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     promoter        complement(4163..4191)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     protein_bind    4386..4509
                     /gene="mutant version of attR"
                     /label=attR4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             complement(4550..4855)
                     /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
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     CDS             complement(5197..5856)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(5910..5940)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     protein_bind    complement(5965..6088)
                     /gene="mutant version of attR"
                     /label=attR3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
ORIGIN
        1 agcttgtcga gaagtactag aggatcataa tcagccatac cacatttgta gaggttttac
       61 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
      121 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
      181 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
      241 atgtatctta tcatgtctgg atctgatcac tgcttgagcc taggagatcc gaaccagata
      301 agtgaaatct agttccaaac tattttgtca tttttaattt tcgtattagc ttacgacgct
      361 acacccagtt cccatctatt ttgtcactct tccctaaata atccttaaaa actccatttc
      421 cacccctccc agttcccaac tattttgtcc gcccacagcg gggcattttt cttcctgtta
      481 tgtttttaat caaacatcct gccaactcca tgtgacaaac cgtcatcttc ggctactttt
      541 tctctgtcac agaatgaaaa tttttctgtc atctcttcgt tattaatgtt tgtaattgac
      601 tgaatatcaa cgcttatttg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca
      661 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
      721 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
      781 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
      841 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
      901 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
      961 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1021 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     1081 ttaacgttta caatttcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt
     1141 ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg
     1201 cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt
     1261 cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta
     1321 aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc
     1381 ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa
     1441 gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc
     1501 cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt
     1561 acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact
     1621 gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac
     1681 aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata
     1741 ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta
     1801 ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg
     1861 gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat
     1921 aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt
     1981 aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga
     2041 aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa
     2101 gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag
     2161 gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac
     2221 tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc
     2281 gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat
     2341 caagagctac caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat
     2401 actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct
     2461 acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt
     2521 cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg
     2581 gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta
     2641 cagcgtgagc attgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg
     2701 gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg
     2761 tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc
     2821 tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg
     2881 gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat
     2941 aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc
     3001 agcgagtcag tgagcgagga agcggaagag cgcctgatgc ggtattttct ccttacgcat
     3061 ctgtgcggta tttcacaccg cagaccagcc gcgtaacctg gcaaaatcgg ttacggttga
     3121 gtaataaatg gatgccctgc gtaagcgggt gtgggcggac aataaagtct taaactgaac
     3181 aaaatagatc taaactatga caataaagtc ttaaactaga cagaatagtt gtaaactgaa
     3241 atcagtccag ttatgctgtg aaaaagcata ctggactttt gttatggcta aagcaaactc
     3301 ttcattttct gaagtgcaaa ttgcccgtcg tattaaagag gggcgtggcc aagggcatgg
     3361 taaagactat attcgcggcg ttgtgacaat ttaccgaaca actccgcggc cgggaagccg
     3421 atctcggctt gaacgaattg ttaggtggcg gtacttgggt cgatatcaaa gtgcatcact
     3481 tcttcccgta tgcccaactt tgtatagaga gccactgcgg gatcgtcacc gtaatctgct
     3541 tgcacgtaga tcacataagc accaagcgcg ttggcctcat gcttgaggag attgatgagc
     3601 gcggtggcaa tgccctgcct ccggtgctcg ccggagactg cgagatcata gatatagatc
     3661 tcactacgcg gctgctcaaa cctgggcaga acgtaagccg cgagagcgcc aacaaccgct
     3721 tcttggtcga aggcagcaag cgcgatgaat gtcttactac ggagcaagtt cccgaggtaa
     3781 tcggagtccg gctgatgttg ggagtaggtg gctacgtctc cgaactcacg accgaaaaga
     3841 tcaagagcag cccgcatgga tttgacttgg tcagggccga gcctacatgt gcgaatgatg
     3901 cccatacttg agccacctaa ctttgtttta gggcgactgc cctgctgcgt aacatcgttg
     3961 ctgctgcgta acatcgttgc tgctccataa catcaaacat cgacccacgg cgtaacgcgc
     4021 ttgctgcttg gatgcccgag gcatagactg tacaaaaaaa cagtcataac aagccatgaa
     4081 aaccgccact gcgccgttac caccgctgcg ttcggtcaag gttctggacc agttgcgtga
     4141 gcgcatacgc tacttgcatt acagtttacg aaccgaacag gcttatgtca actgggttcg
     4201 tgccttcatc cgtttccacg gtgtgcgtca cccggcaacc ttgggcagca gcgaagtcga
     4261 ggcatttctg tcctggctgg cgaacgagcg caaggtttcg gtctccacgc atcgtcaggc
     4321 attggcggcc ttgctgttct tctacggcaa ggtgctgtgc acggatctac catgggatcc
     4381 tcgatcaact ttgtatagaa aagttgaacg agaaacgtaa aatgatataa atatcaatat
     4441 attaaattag attttgcata aaaaacagac tacataatac tgtaaaacac aacatatcca
     4501 gtcactatgg tcgacctgca gactggctgt gtataaggga gcctgacatt tatattcccc
     4561 agaacatcag gttaatggcg tttttgatgt cattttcgcg gtggctgaga tcagccactt
     4621 cttccccgat aacggagacc ggcacactgg ccatatcggt ggtcatcatg cgccagcttt
     4681 catccccgat atgcaccacc gggtaaagtt cacgggggac tttatctgac agcagacgtg
     4741 cactggccag ggggatcacc atccgtcgcc cgggcgtgtc aataatatca ctctgtacat
     4801 ccacaaacag acgataacgg ctctctcttt tataggtgta aaccttaaac tgcatttcac
     4861 cagcccctgt tctcgtcggc aaaagagccg ttcatttcaa taaaccgggc gacctcagcc
     4921 atcccttcct gattttccgc tttccagcgt tcggcacgca gacgacgggc ttcattctgc
     4981 atggttgtgc ttaccgaacc ggagatattg acatcatata tgccttgagc aactgatagc
     5041 tgtcgctgtc aactgtcact gtaatacgct gcttcatagc atacctcttt ttgacatact
     5101 tcgggtatac atatcagtat atattcttat accgcaaaaa tcagcgcgca aatacgcata
     5161 ctgttatctg gcttttagta agccggatcc tctagattac gccccgccct gccactcatc
     5221 gcagtactgt tgtaattcat taagcattct gccgacatgg aagccatcac aaacggcatg
     5281 atgaacctga atcgccagcg gcatcagcac cttgtcgcct tgcgtataat atttgcccat
     5341 ggtgaaaacg ggggcgaaga agttgtccat attggccacg tttaaatcaa aactggtgaa
     5401 actcacccag ggattggctg agacgaaaaa catattctca ataaaccctt tagggaaata
     5461 ggccaggttt tcaccgtaac acgccacatc ttgcgaatat atgtgtagaa actgccggaa
     5521 atcgtcgtgg tattcactcc agagcgatga aaacgtttca gtttgctcat ggaaaacggt
     5581 gtaacaaggg tgaacactat cccatatcac cagctcaccg tctttcattg ccatacggaa
     5641 ttccggatga gcattcatca ggcgggcaag aatgtgaata aaggccggat aaaacttgtg
     5701 cttatttttc tttacggtct ttaaaaaggc cgtaatatcc agctgaacgg tctggttata
     5761 ggtacattga gcaactgact gaaatgcctc aaaatgttct ttacgatgcc attgggatat
     5821 atcaacggtg gtatatccag tgattttttt ctccatttta gcttccttag ctcctgaaaa
     5881 tctcgacgga tcctaactca aaatccacac attatacgag ccggaagcat aaagtgtaaa
     5941 gcctggggtg cctaatgcgg ccgccatagt gactggatat gttgtgtttt acagtattat
     6001 gtagtctgtt ttttatgcaa aatctaattt aatatattga tatttatatc attttacgtt
     6061 tctcgttcaa ctttattata catagttgat cgagctagca
//
