LOCUS       Exported                6969 bp ds-DNA     circular SYN 15-NOV-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 6969)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 11, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6969
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     rep_origin      129..584
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     polyA_signal    715..763
                     /note="synthetic polyadenylation signal"
     misc_feature    777..868
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     869..897
                     
     enhancer        896..2963
                     /label= hs GLAST promotor
                     /note="GenBank: AF448436.1"
     primer_bind     complement(919..941)
                     /label=oJDW 756 GLAST 2
     primer_bind     2654..2673
                     /label=GLAST FWD
     primer_bind     2935..2990
                     /label=oJDW 1676
     primer_bind     complement(2945..2964)
                     /label=oJDW 1596 hsGLAST REV +  AgeI
     promoter        2964..3011
                     /label=CMV minimal pro
     protein_bind    3023..3147
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        3172..3202
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     CDS             3256..3915
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     3284..3308
                     /label=oJDW 1397
     primer_bind     complement(3322..3341)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     complement(3427..3450)
                     /label=oJDW 1402
     primer_bind     3848..3868
                     /label=oJDW 1396
     CDS             4257..4562
                     /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"
     primer_bind     4441..4460
                     /label=ccdB-fwd
                     /note="ccdB gene, forward primer"
     protein_bind    complement(4603..4727)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4742..4792)
                     /label=oJDW 1677
     polyA_signal    4809..4930
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4874..4893
                     /label=SV40pA REVSeq
     rep_origin      complement(5349..5939)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6109..6969)
                     /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"
     primer_bind     complement(6197..6216)
                     /label=oJDW 856
     primer_bind     6933..6952
                     /label=oJDW 840
ORIGIN
        1 actntccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
       61 atatttgaat gtattagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
      121 tgccacngac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
      181 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
      241 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
      301 ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg
      361 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
      421 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt
      481 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
      541 aaaatttaac gcgaatttta acaaaatatt aacgcttaca atttgccatt cgccattcag
      601 gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagcccaa
      661 gctaccatga taagtaagta atattaaggt acgggaggta cttggagcgg ccgcaataaa
      721 atatctttat tttcattaca tctgtgtgtt ggttttttgt gtgaatcgat agtactaaca
      781 tacgctctcc atcaaaacaa aacgaaacaa aacaaactag caaaataggc tgtccccagt
      841 gcaagtgcag gtgccagaac atttctctat cgataggtac catgtctaca caaactgaaa
      901 taatgaaatg gtttttaacc aagctttagg gattagagag gaaaaaaaaa gtgaaaaaga
      961 aagggcacca gggtcttccc atctctatgc taaccaaagt gtggttttcc agcccttttt
     1021 cttgcctatt aacaaaaata aaaacaaaaa atggaaatgc cactgggctt gaaaccccat
     1081 ttaaaatcag atttctcttt cgagttccct tgcaatacaa gtcaaagtca tttccaccaa
     1141 aaaaaaaaag cggtgggggg gggtgcgcaa attaatccta aactcatggc tcagatagga
     1201 cctggtatag taaataaatg cttttaaaag tagtcatggc attgagaaat ggctccatgc
     1261 acctaaacca gaggcaccaa tgtaaaaact agtaattgtg ttttaacaca aaagattgaa
     1321 aagcgttttg tttcacatcc taatattatt ccacccactt aaaaagagtt acttaatatt
     1381 gactctgtat taggtataag acaagttctt gccctcaaag aacttataat ttagttgaga
     1441 aaccaaaact taacatgagc ttaaaaagaa ggatgcagta caaaacatgg tataattaag
     1501 tgctaaacaa tgtagtatag atgctcagtg acctaggaga tcagaggaag ggaagggtca
     1561 ctgagggctg gggatgtcaa aagcatgtac tttcagtaag ttcttaaaat gatagcatgt
     1621 tagaaagctg ttataattgc tgctgatttg taattttaaa gcgttgacat gattcaccat
     1681 tgctttccaa ccagtagctg tttaaaagga catacagttg agggtgaaag gagatatttc
     1741 tttatgatct ccagagattt gtattgaggg gaaaaaagac atactttctt gttgaataat
     1801 agaccaagaa ttgagcatat tttgcatggg gaatattaaa ttcagtgaac aagaatccaa
     1861 acctcaggct gtgattctgg tataaatttt tgtttctaat gaaccatttt cttccttcta
     1921 aagaagattt aagttttggg aataagtcac ctcaaaaaat ttttttttta ttttctaata
     1981 tctattttca caatttattt ttdttttaag aaaagtggac tagaagctca agttaccttc
     2041 tcaaggcaat gagaattttt tagctctaaa aaagttgtgt gtggtaggga gaacatttag
     2101 catcctttaa aatggagaca tctgaacagc aacctatctt ttgaatcgcc aaatctcaag
     2161 tgtactttgc actctcggtc ttaaaatgct cttcattttc tagtttggca aaaattccag
     2221 tctctttaca tttgctacct tttccgtctt tgcaacacag gcagtgtgag ctaaattagg
     2281 tttgttaaaa tgaagttttg aagtttcaac tgctgaaata gaggcatgtc cctaacttta
     2341 gacaaattta gagaatcgta gtaagcatta ttttttttct acatacttgt ctgcattcag
     2401 tgtcttccca agaaacgtca gttaattccc cgattcgaaa aaggaacttt ctcgtaacag
     2461 ttgtacaacc aggctaaaga gcacaccctc gtcttccctg aaagggagga aacatgccat
     2521 aatgtgaggt aattaaattt agccttagaa aagtctttga aacgttctaa tcaagacttt
     2581 cagctttccg attgtgtgcg ctttattttt caattcttcc tagtcccggt tttgcatttc
     2641 taatgacagt gaactgacaa agcgtgaaag tggtctaagg agcaaaacaa agccagcaag
     2701 cctgcctctg gtgtgccaag attaatcaat attccagaaa cctcggggtt tccccctcct
     2761 ccctgtgcgg tttccattcc ccactcctcc tttcccgggc acagactcct ccccttttct
     2821 taggttgccc tgatagtaac ttgcggtttc agagcacatg cacactgtca gggctagcct
     2881 gcctgcttac gcgcgctgcg gattgctgct ccgttgtacc tgctggggaa ttcacctcgt
     2941 tactgcttga tgggctcgag atcctgtttt gacctccata gaagacaccg ggaccgatcc
     3001 agcctccgcg gtggcggcca tcacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa
     3061 tgatataaat atcaatatat taaattagat tttgcataaa aaacagacta cataatactg
     3121 taaaacacaa catatccagt cactatggcg gccgcattag gcaccccagg ctttacactt
     3181 tatgcttccg gctcgtataa tgtgtggatt ttgagttagg atccgtcgag attttcagga
     3241 gctaaggaag ctaaaatgga gaaaaaaatc actggatata ccaccgttga tatatcccaa
     3301 tggcatcgta aagaacattt tgaggcattt cagtcagttg ctcaatgtac ctataaccag
     3361 accgttcagc tggatattac ggccttttta aagaccgtaa agaaaaataa gcacaagttt
     3421 tatccggcct ttattcacat tcttgcccgc ctgatgaatg ctcatccgga attccgtatg
     3481 gcaatgaaag acggtgagct ggtgatatgg gatagtgttc acccttgtta caccgttttc
     3541 catgagcaaa ctgaaacgtt ttcatcgctc tggagtgaat accacgacga tttccggcag
     3601 tttctacaca tatattcgca agatgtggcg tgttacggtg aaaacctggc ctatttccct
     3661 aaagggttta ttgagaatat gtttttcgtc tcagccaatc cctgggtgag tttcaccagt
     3721 tttgatttaa acgtggccaa tatggacaac ttcttcgccc ccgttttcac catgggcaaa
     3781 tattatacgc aaggcgacaa ggtgctgatg ccgctggcga ttcaggttca tcatgccgtt
     3841 tgtgatggct tccatgtcgg cagaatgctt aatgaattac aacagtactg cgatgagtgg
     3901 cagggcgggg cgtaaacgcg tggatccggc ttactaaaag ccagataaca gtatgcgtat
     3961 ttgcgcgctg atttttgcgg tataagaata tatactgata tgtatacccg aagtatgtca
     4021 aaaagaggta tgctatgaag cagcgtatta cagtgacagt tgacagcgac agctatcagt
     4081 tgctcaaggc atatatgatg tcaatatctc cggtctggta agcacaacca tgcagaatga
     4141 agcccgtcgt ctgcgtgccg aacgctggaa agcggaaaat caggaaggga tggctgaggt
     4201 cgcccggttt attgaaatga acggctcttt tgctgacgag aacaggggct ggtgaaatgc
     4261 agtttaaggt ttacacctat aaaagagaga gccgttatcg tctgtttgtg gatgtacaga
     4321 gtgatattat tgacacgccc gggcgacgga tggtgatccc cctggccagt gcacgtctgc
     4381 tgtcagataa agtctcccgt gaactttacc cggtggtgca tatcggggat gaaagctggc
     4441 gcatgatgac caccgatatg gccagtgtgc cggtctccgt tatcggggaa gaagtggctg
     4501 atctcagcca ccgcgaaaat gacatcaaaa acgccattaa cctgatgttc tggggaatat
     4561 aaatgtcagg ctcccttata cacagccagt ctgcaggtcg accatagtga ctggatatgt
     4621 tgtgttttac agtattatgt agtctgtttt ttatgcaaaa tctaatttaa tatattgata
     4681 tttatatcat tttacgtttc tcgttcagct ttcttgtaca aagtggtgat ggccgctcta
     4741 gcgaagggat cccccatcga atgcatgcat ctagagtcgg ggcggccggc cgcttcgagc
     4801 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     4861 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     4921 taaacaagtt aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg
     4981 ggaggttttt taaagcaagt aaaacctcta caatgtggta aaatcgatna ggatccgtcg
     5041 accgatgccc ttgagagcct tcaacccagt cagctccttc cggtgggcgc ggggcatgac
     5101 tatcgtcgcc gcacttatga ctgtcttctt tatcatgcaa ctcgtangac aggtgccggc
     5161 agcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag
     5221 cggtatcagc tcactcagag gcggtaatac ggttatccac agaatcangg gataacgcan
     5281 gaagaacatg tgagcaaaan gccagcaaaa ggccaggaaa ccgtanaaat ggccgcgttg
     5341 ctggcgtttt tccataggct ccgccccccc tgacgagcat cacaaaaatc gacgctcaag
     5401 tcagaggtgg cgaaacccga cagggactat aaagatacca ggcgtttccc cctggaagct
     5461 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttacngg atacctgtcc gcctttctcc
     5521 cttcgggaag cgtggcgctt tctcatagct cacgcngtag gtatctcagt tcggtgtagg
     5581 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     5641 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgactnatcg ccactggcag
     5701 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     5761 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     5821 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5881 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5941 aagatccttt gatctttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     6001 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     6061 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     6121 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     6181 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     6241 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     6301 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     6361 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     6421 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     6481 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     6541 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     6601 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     6661 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     6721 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     6781 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     6841 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     6901 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     6961 aatactcat
//
