LOCUS       Exported                6223 bp ds-DNA     circular UNA 23-MAR-2022
DEFINITION  natural circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6223)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Mar 23, 2022 from SnapGene 4.0.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6223
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     promoter        61..105
                     /label=CMVd3 promoter
                     /note="truncated human cytomegalovirus (CMV) immediate
                     early promoter that yields strongly reduced expression 
                     relative to the full-length promoter"
     misc_feature    150..227
                     /label=IRE1 signal peptide
                     /label=Lumenal
                     /note="/ApEinfo_fwdcolor=#b7e6d7
                     /ApEinfo_revcolor=#b7e6d7"
     misc_feature    228..290
                     /label=TM
     CDS             306..335
                     /codon_start=1
                     /label=GS 10x v1
                     /translation="GGSGGGGSGG"
     CDS             336..989
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     CDS             990..1016
                     /codon_start=1
                     /label=GS
                     /label=Translation 474-1520
                     /translation="GSGGGGSGG"
     CDS             1017..1906
                     /codon_start=1
                     /label=HaloTag Core
                     /note="HaloTag Core"
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEIS"
     CDS             1920..1940
                     /codon_start=1
                     /label=Linker Peptide
                     /note="Linker Peptide"
                     /translation="YKSGLRS"
     CDS             1941..1970
                     /codon_start=1
                     /label=10x GS # 4
                     /translation="GGGGSGGGGS"
     CDS             1977..2867
                     /codon_start=1
                     /label=HaloTag Core
                     /note="HaloTag Core"
                     /translation="MAEIGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSS
                     YVWRNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVI
                     HDWGSALGFHWAKRNPERVKGIAFMEFIRPIPTWDEWPEFARETFQAFRTTDVGRKLII
                     DQNVFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALV
                     EEYMDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDL
                     IGSEIARWLSTLEISG"
     misc_feature    2868..2879
                     /label=Retention signal
     CDS             2880..2882
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=STOP
                     /translation=""
     polyA_signal    3011..3132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3139..3594)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3621..3725
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3727..4084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3935..4070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4119..4913
                     /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
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5145..5192
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5521..6109
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagctggttt agtgaaccgt
      121 cagatccgct agcgctaccg gtcgccacca tgccggcccg gcggctgctg ctgctgctga
      181 cgctgctgct gcccggcctc gggatttttg gaagtatgct taaggacatg gctaccatca
      241 tcctgagcac cttcctgctg attggctggg tggccttcat catcacctat cccctgagca
      301 tgcatggagg atctggaggc ggcgggtccg gcggaatgtc ccctatacta ggttattgga
      361 aaattaaggg ccttgtgcaa cccactcgac ttcttttgga atatcttgaa gaaaaatatg
      421 aagagcattt gtatgagcgc gatgaaggtg ataaatggcg aaacaaaaag tttgaattgg
      481 gtttggagtt tcccaatctt ccttattata ttgatggtga tgttaaatta acacagtcta
      541 tggccatcat acgttatata gctgacaagc acaacatgtt gggtggttgt ccaaaagagc
      601 gtgcagagat ttcaatgctt gaaggagcgg ttttggatat tagatacggt gtttcgagaa
      661 ttgcatatag taaagacttt gaaactctca aagttgattt tcttagcaag ctacctgaaa
      721 tgctgaaaat gttcgaagat cgtttatgtc ataaaacata tttaaatggt gatcatgtaa
      781 cccatcctga cttcatgttg tatgacgctc ttgatgttgt tttatacatg gacccaatgt
      841 gcctggatgc gttcccaaaa ttagtttgtt ttaaaaaacg tattgaagct atcccacaaa
      901 ttgataagta cttgaaatcc agcaagtata tagcatggcc tttgcagggc tggcaagcca
      961 cgtttggtgg tggcgaccat cctccaaaag gatctggagg cggcgggtcc ggcggaatgg
     1021 cagaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca
     1081 tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc
     1141 cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca
     1201 ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg
     1261 acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc
     1321 tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc
     1381 gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc
     1441 cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga
     1501 tcatcgatca gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga
     1561 ctgaagtcga gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac
     1621 tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg
     1681 tcgaagaata catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca
     1741 ccccaggcgt tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact
     1801 gcaaggctgt ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga
     1861 tcggcagcga gatcgcgcgc tggctgtcga cgctcgagat ttccggcatg gacgagctgt
     1921 acaagtccgg actcagatct ggtggaggcg gtagcggagg aggtggctct gtcgacatgg
     1981 cagaaatcgg tactggcttt ccattcgacc cccattatgt ggaagtcctg ggcgagcgca
     2041 tgcactacgt cgatgttggt ccgcgcgatg gcacccctgt gctgttcctg cacggtaacc
     2101 cgacctcctc ctacgtgtgg cgcaacatca tcccgcatgt tgcaccgacc catcgctgca
     2161 ttgctccaga cctgatcggt atgggcaaat ccgacaaacc agacctgggt tatttcttcg
     2221 acgaccacgt ccgcttcatg gatgccttca tcgaagccct gggtctggaa gaggtcgtcc
     2281 tggtcattca cgactggggc tccgctctgg gtttccactg ggccaagcgc aatccagagc
     2341 gcgtcaaagg tattgcattt atggagttca tccgccctat cccgacctgg gacgaatggc
     2401 cagaatttgc ccgcgagacc ttccaggcct tccgcaccac cgacgtcggc cgcaagctga
     2461 tcatcgatca gaacgttttt atcgagggta cgctgccgat gggtgtcgtc cgcccgctga
     2521 ctgaagtcga gatggaccat taccgcgagc cgttcctgaa tcctgttgac cgcgagccac
     2581 tgtggcgctt cccaaacgag ctgccaatcg ccggtgagcc agcgaacatc gtcgcgctgg
     2641 tcgaagaata catggactgg ctgcaccagt cccctgtccc gaagctgctg ttctggggca
     2701 ccccaggcgt tctgatccca ccggccgaag ccgctcgcct ggccaaaagc ctgcctaact
     2761 gcaaggctgt ggacatcggc ccgggtctga atctgctgca agaagacaac ccggacctga
     2821 tcggcagcga gatcgcgcgc tggctgtcga cgctcgagat ttccggcaaa aagatgccat
     2881 aagatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     2941 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3001 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3061 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3121 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3181 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3241 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3301 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3361 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3421 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3481 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3541 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3601 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     3661 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     3721 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3781 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3901 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3961 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4021 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4081 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4141 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4201 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4261 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4321 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4381 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4441 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4501 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4561 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4621 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     4681 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4741 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4801 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4861 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4921 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4981 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5041 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5101 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5161 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5221 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5281 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5341 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5401 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5461 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5521 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5581 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     5641 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt
     5701 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     5761 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     5821 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     5881 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     5941 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6001 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6061 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6121 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6181 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
