LOCUS       Exported                7374 bp ds-DNA     circular SYN 18-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7374)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 2020 from SnapGene 4.1.9
            http://www.snapgene.com
COMMENT     Ligation of : pAC8HGW.txt [7291 nt] : (EcoRV-Blunt(filled)[5274] / 
            EcoRV[5274]) to LoxP.txt.xdna [34 nt] : (Uncut 5'[0] / Uncut 3'[34])
FEATURES             Location/Qualifiers
     source          1..7374
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      1422..1877
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2159..2263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             2264..3124
                     /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      3295..3883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     misc_recomb     4446..5273
                     /label=baculovirus recombination region (lef2/ORF603)
                     /note="contains ORF603 and part of lef2"
     CDS             4446..4570
                     /codon_start=3
                     /product="baculovirus late expression factor 2"
                     /label=lef2
                     /translation="HLLIKSQDVYKPPNCQKMKTVDKLCPFAGNCKGLNPICNY"
     CDS             complement(4608..5213)
                     /codon_start=1
                     /product="baculovirus ORF603 protein"
                     /label=ORF603
                     /translation="MAVIFNNKQLLADNSIEKGGELFLFNGSYNILESYVNPVLLKNGV
                     IELEEAAYYAGNILYKTDDPKFIDYINLIIKATHSEELPENSTVVNYRKTMRSGTIHPI
                     KKDIYIYDNKKFTLYDRYIYGYDNNYVNFYEEKNEKEKEYEEEDDKASSLCENKIILSQ
                     INCESFENDFKYYLSDYNYAFSIIDNTTNVLVAFGLYR"
     protein_bind    5277..5310
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        5311..5402
                     /gene="polh from Autographa californica"
                     /label=polyhedrin promoter
                     /note="promoter for the baculovirus polyhedrin gene"
     primer_bind     complement(5335..5354)
                     /label=Bac1_RV
     CDS             5426..5455
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             5465..5488
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     protein_bind    5513..5637
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        5662..5692
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             5746..6405
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     CDS             6747..7052
                     /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(7093..7217)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     CDS             7225..7254
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     misc_recomb     join(7292..7374,1..1275)
                     /label=baculovirus recombination region (ORF1629)
                     /note="contains part of ORF1629"
     CDS             complement(join(7308..7374,1..1275))
                     /codon_start=2
                     /product="baculovirus capsid-associated protein"
                     /label=ORF1629
                     /note="required for viral replication"
                     /translation="PSLRQSLYNTIAYIERLLNIGTVNDSEITMLIADFYDLYSNYNIE
                     LPPPQALPRSRRPSVVQPAAPAPVPTIVREQTKPEQIIPAAPPPPPSPVPNIPAPPPPP
                     PPSMSELPPAPPMPTEPQPAAPLDDRQQLLEAIRNEKNRTRLRPVKPKTAPETSTIVEV
                     PTVLPKETFEPKPPSASPPPPPPPPPPPAPPAPPPMVDLSSAPPPPPLVDLPSEMLPPP
                     APSLSNVLSELKSGTVRLKPAQKRPQSEIIPKSSTTNLIADVLADTINRRRVAMAKSSS
                     EATSNDEGWDDDDNRPNKANTPDVKYVQALFNVFTSSQLYTNDSDERNTKAHNILNDVE
                     PLLQNKTQTNIDKARLLLQDLASFVALSENPLDSPAIGSEKQPLFETNRNLFYKSIEDL
                     IFKFRYKDAENHLIFALTYHPKDYKFNELLKYVQQLSVNQQRTESSA"
ORIGIN
        1 aatttataat ctttagggtg gtatgttaga gcgaaaatca aatgattttc agcgtcttta
       61 tatctgaatt taaatattaa atcctcaata gatttgtaaa ataggtttcg attagtttca
      121 aacaagggtt gtttttccga accgatggct ggactatcta atggattttc gctcaacgcc
      181 acaaaacttg ccaaatcttg tagcagcaat ctagctttgt cgatattcgt ttgtgttttg
      241 ttttgtaata aaggttcgac gtcgttcaaa atattatgcg cttttgtatt tctttcatca
      301 ctgtcgttag tgtacaattg actcgacgta aacacgttaa ataaagcttg gacatattta
      361 acatcgggcg tgttagcttt attaggccga ttatcgtcgt cgtcccaacc ctcgtcgtta
      421 gaagttgctt ccgaagacga ttttgccata gccacacgac gcctattaat tgtgtcggct
      481 aacacgtccg cgatcaaatt tgtagttgag ctttttggaa ttatttctga ttgcgggcgt
      541 ttttgggcgg gtttcaatct aactgtgccc gattttaatt cagacaacac gttagaaagc
      601 gatggtgcag gcggtggtaa catttcagac ggcaaatcta ctaatggcgg cggtggtgga
      661 gctgatgata aatctaccat cggtggaggc gcaggcgggg ctggcggcgg aggcggaggc
      721 ggaggtggtg gcggtgatgc agacggcggt ttaggctcaa atgtctcttt aggcaacaca
      781 gtcggcacct caactattgt actggtttcg ggcgccgttt ttggtttgac cggtctgaga
      841 cgagtgcgat ttttttcgtt tctaatagct tccaacaatt gttgtctgtc gtctaaaggt
      901 gcagcgggtt gaggttccgt cggcattggt ggagcgggcg gcaattcaga catcgatggt
      961 ggtggtggtg gtggaggcgc tggaatgtta ggcacgggag aaggtggtgg cggcggtgcc
     1021 gccggtataa tttgttctgg tttagtttgt tcgcgcacga ttgtgggcac cggcgcaggc
     1081 gccgctggct gcacaacgga aggtcgtctg cttcgaggca gcgcttgggg tggtggcaat
     1141 tcaatattat aattggaata caaatcgtaa aaatctgcta taagcattgt aatttcgcta
     1201 tcgtttaccg tgccgatatt taacaaccgc tcaatgtaag caattgtatt gtaaagagat
     1261 tgtctcaagc tcggatcgat cccgcacgcc gataacaagc cttttcattt ttactacagc
     1321 attgtagtgg cgagacactt cgctgtcgtc gcctgatgcg gtattttctc cttacgcatc
     1381 tgtgcggtat ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc
     1441 attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgccct
     1501 agcgcccgct cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg
     1561 tcaagctcta aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga
     1621 ccccaaaaaa cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt
     1681 ttttcgccct ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg
     1741 aacaacactc aaccctatct cgggctattc ttttgattta taagggattt tgccgatttc
     1801 ggcctattgg ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat
     1861 attaacgttt acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt
     1921 aagccagccc cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc
     1981 ggcatccgct tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc
     2041 accgtcatca ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt
     2101 taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg
     2161 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     2221 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     2281 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     2341 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     2401 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     2461 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     2521 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     2581 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     2641 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     2701 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     2761 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     2821 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2881 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2941 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     3001 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     3061 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     3121 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     3181 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     3241 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     3301 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     3361 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     3421 agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc acttcaagaa
     3481 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     3541 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     3601 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     3661 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     3721 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     3781 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3841 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3901 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3961 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     4021 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     4081 accgcctctc cccgcgcgtt ggccgattca ttaatgcaga acggctccgc ccactattaa
     4141 tgaaattaaa aattccaatt ttaaaaaacg cagcaagaga aacatttgta tgaaagaatg
     4201 cgtagaagga aagaaaaatg tcgtcgacat gctgaacaac aagattaata tgcctccgtg
     4261 tataaaaaaa atattgaacg atttgaaaga aaacaatgta ccgcgcggcg gtatgtacag
     4321 gaagaggttt atactaaact gttacattgc aaacgtggtt tcgtgtgcca agtgtgaaaa
     4381 ccgatgttta atcaaggctc tgacgcattt ctacaaccac gactccaagt gtgtgggtga
     4441 agtcatgcat cttttaatca aatcccaaga tgtgtataaa ccaccaaact gccaaaaaat
     4501 gaaaactgtc gacaagctct gtccgtttgc tggcaactgc aagggtctca atcctatttg
     4561 taattattga ataataaaac aattataaat gctaaatttg ttttttatta acgatacaaa
     4621 ccaaacgcaa caagaacatt tgtagtatta tctataattg aaaacgcgta gttataatcg
     4681 ctgaggtaat atttaaaatc attttcaaat gattcacagt taatttgcga caatataatt
     4741 ttattttcac ataaactaga cgccttgtcg tcttcttctt cgtattcctt ctctttttca
     4801 tttttctcct cataaaaatt aacatagtta ttatcgtatc catatatgta tctatcgtat
     4861 agagtaaatt ttttgttgtc ataaatatat atgtcttttt taatggggtg tatagtaccg
     4921 ctgcgcatag tttttctgta atttacaaca gtgctatttt ctggtagttc ttcggagtgt
     4981 gttgctttaa ttattaaatt tatataatca atgaatttgg gatcgtcggt tttgtacaat
     5041 atgttgccgg catagtacgc agcttcttct agttcaatta caccattttt tagcagcacc
     5101 ggattaacat aactttccaa aatgttgtac gaaccgttaa acaaaaacag ttcacctccc
     5161 ttttctatac tattgtctgc gagcagttgt ttgttgttaa aaataacagc cattgtaatg
     5221 agacgcacaa actaatatca caaactggaa atgtctatca atatatagtt gctgatataa
     5281 cttcgtatag catacattat acgaagttat atcatggaga taattaaaat gataaccatc
     5341 tcgcaaataa ataagtattt tactgttttc gtaacagttt tgtaataaaa aaacctataa
     5401 atacggatct accatgggca gcagccacca tcaccatcat catcatcatc atcactccgc
     5461 gggtctggaa gttctgttcc aggggcccca tatgggatcc gaattcccat caacaagttt
     5521 gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat taaattagat
     5581 tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt cactatggcg
     5641 gccgcattag gcaccccagg ctttacactt tatgcttccg gctcgtataa tgtgtggatt
     5701 ttgagttagg atccgtcgag attttcagga gctaaggaag ctaaaatgga gaaaaaaatc
     5761 actggatata ccaccgttga tatatcccaa tggcatcgta aagaacattt tgaggcattt
     5821 cagtcagttg ctcaatgtac ctataaccag accgttcagc tggatattac ggccttttta
     5881 aagaccgtaa agaaaaataa gcacaagttt tatccggcct ttattcacat tcttgcccgc
     5941 ctgatgaatg ctcatccgga attccgtatg gcaatgaaag acggtgagct ggtgatatgg
     6001 gatagtgttc acccttgtta caccgttttc catgagcaaa ctgaaacgtt ttcatcgctc
     6061 tggagtgaat accacgacga tttccggcag tttctacaca tatattcgca agatgtggcg
     6121 tgttacggtg aaaacctggc ctatttccct aaagggttta ttgagaatat gtttttcgtc
     6181 tcagccaatc cctgggtgag tttcaccagt tttgatttaa acgtggccaa tatggacaac
     6241 ttcttcgccc ccgttttcac catgggcaaa tattatacgc aaggcgacaa ggtgctgatg
     6301 ccgctggcga ttcaggttca tcatgccgtt tgtgatggct tccatgtcgg cagaatgctt
     6361 aatgaattac aacagtactg cgatgagtgg cagggcgggg cgtaaacgcg tggatccggc
     6421 ttactaaaag ccagataaca gtatgcgtat ttgcgcgctg atttttgcgg tataagaata
     6481 tatactgata tgtatacccg aagtatgtca aaaagaggta tgctatgaag cagcgtatta
     6541 cagtgacagt tgacagcgac agctatcagt tgctcaaggc atatatgatg tcaatatctc
     6601 cggtctggta agcacaacca tgcagaatga agcccgtcgt ctgcgtgccg aacgctggaa
     6661 agcggaaaat caggaaggga tggctgaggt cgcccggttt attgaaatga acggctcttt
     6721 tgctgacgag aacaggggct ggtgaaatgc agtttaaggt ttacacctat aaaagagaga
     6781 gccgttatcg tctgtttgtg gatgtacaga gtgatattat tgacacgccc gggcgacgga
     6841 tggtgatccc cctggccagt gcacgtctgc tgtcagataa agtctcccgt gaactttacc
     6901 cggtggtgca tatcggggat gaaagctggc gcatgatgac caccgatatg gccagtgtgc
     6961 cggtctccgt tatcggggaa gaagtggctg atctcagcca ccgcgaaaat gacatcaaaa
     7021 acgccattaa cctgatgttc tggggaatat aaatgtcagg ctcccttata cacagccagt
     7081 ctgcaggtcg accatagtga ctggatatgt tgtgttttac agtattatgt agtctgtttt
     7141 ttatgcaaaa tctaatttaa tatattgata tttatatcat tttacgtttc tcgttcagct
     7201 ttcttgtaca aagtggtagg atccgaacaa aaactcatct cagaagagga tctgtaagct
     7261 tgatgggcgg ccgcttaatt aattgatccg ggttattagt acatttatta agcgctagat
     7321 tctgtgcgtt gttgatttac agacaattgt tgtacgtatt ttaataattc atta
//
