LOCUS       Exported                5572 bp ds-DNA     circular SYN 03-MAY-2023
DEFINITION  An improved universal plasmid for expression of circRNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5572)
  AUTHORS   Brett Stringer
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jul 20, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Created by Brett Stringer, Flinders University, 18 July 2022.
FEATURES             Location/Qualifiers
     source          1..5572
                     /organism="synthetic DNA construct"
                     /lab_host="Escherichia coli"
                     /mol_type="other DNA"
     primer_bind     133..152
                     /label=BWS533
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     836..855
                     /label=BWS044
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     926..945
                     /label=BWS335
     primer_bind     947..966
                     /label=BWS409
     intron          958..1044
                     /label=ZKSCAN1 intron 1 partial
     misc_feature    958..997
                     /label=AluSq2
     misc_feature    1027..1044
                     /label=splice site
     misc_feature    1046..1051
                     /label=BsmBI
     misc_feature    1060..1065
                     /label=BsmBI
     intron          1067..1125
                     /label=ZKSCAN1 intron 3 partial
     misc_feature    1067..1072
                     /label=splice site
     misc_feature    1090..1125
                     /label=AluSz
     polyA_signal    1169..1393
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1174..1193)
                     /label=BWS337
     primer_bind     complement(1174..1193)
                     /label=SC4001
     primer_bind     complement(1276..1296)
                     /label=SC4000
     rep_origin      1439..1867
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        1881..2210
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2061..2196
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2277..3071
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3245..3366
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3415..3431)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    3439..3455
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /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)."
     promoter        complement(3463..3493)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3508..3529
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(3817..4405)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4576..5436)
                     /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     5150..5169
                     /label=BWS534
     promoter        complement(5437..5541)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattcaaa
      961 gtgctgagat tacaggcgtg agccaccacc cccggcccac tttttgtaaa ggtacgtact
     1021 aatgactttt tttttatact tcagggagac ggttgtgtac gtctcggtaa gaagcaagga
     1081 aaagaattag gctcggcacg gtagctcaca cctgtaatcc cagcactcga gtctagaggg
     1141 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     1201 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     1261 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     1321 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     1381 gtgggctcta tggcttctga ggcggaaaga accagctggg gctctagggg gtatccccac
     1441 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct
     1501 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg
     1561 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt
     1621 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca
     1681 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga
     1741 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa
     1801 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac
     1861 gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag
     1921 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     1981 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     2041 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     2101 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     2161 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     2221 gagcttgtat atccattttc ggatctgatc aagagacagg atgaggatcg tttcgcatga
     2281 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     2341 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     2401 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcagg
     2461 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     2521 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     2581 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     2641 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     2701 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     2761 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgcgcatg cccgacggcg
     2821 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     2881 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     2941 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     3001 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     3061 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     3121 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     3181 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     3241 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     3301 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     3361 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     3421 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     3481 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     3541 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     3601 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     3661 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     3721 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     3781 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     3841 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     3901 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     3961 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     4021 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     4081 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     4141 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     4201 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac
     4261 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     4321 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     4381 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     4441 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     4501 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     4561 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     4621 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     4681 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     4741 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     4801 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     4861 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     4921 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     4981 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     5041 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     5101 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     5161 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     5221 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     5281 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     5341 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     5401 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     5461 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     5521 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tc
//
