LOCUS       Exported                8820 bp ds-DNA     circular SYN 14-SEP-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8820)
  AUTHORS   Yan Cui
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Aug 6, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8820
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             890..913
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             932..4354
                     /codon_start=1
                     /label=DDB1
                     /translation="MSYNYVVTAQKPTAVNGCVTGHFTSAEDLNLLIAKNTRLEIYVVT
                     AEGLRPVKEVGMYGKIAVMELFRPKGESKDLLFILTAKYNACILEYKQSGESIDIITRA
                     HGNVQDRIGRPSETGIIGIIDPECRMIGLRLYDGLFKVIPLDRDNKELKAFNIRLEELH
                     VIDVKFLYGCQAPTICFVYQDPQGRHVKTYEVSLREKEFNKGPWKQENVEAEASMVIAV
                     PEPFGGAIIIGQESITYHNGDKYLAIAPPIIKQSTIVCHNRVDPNGSRYLLGDMEGRLF
                     MLLLEKEEQMDGTVTLKDLRVELLGETSIAECLTYLDNGVVFVGSRLGDSQLVKLNVDS
                     NEQGSYVVAMETFTNLGPIVDMCVVDLERQGQGQLVTCSGAFKEGSLRIIRNGIGIHEH
                     ASIDLPGIKGLWPLRSDPNRETDDTLVLSFVGQTRVLMLNGEEVEETELMGFVDDQQTF
                     FCGNVAHQQLIQITSASVRLVSQEPKALVSEWKEPQAKNISVASCNSSQVVVAVGRALY
                     YLQIHPQELRQISHTEMEHEVACLDITPLGDSNGLSPLCAIGLWTDISARILKLPSFEL
                     LHKEMLGGEIIPRSILMTTFESSHYLLCALGDGALFYFGLNIETGLLSDRKKVTLGTQP
                     TVLRTFRSLSTTNVFACSDRPTVIYSSNHKLVFSNVNLKEVNYMCPLNSDGYPDSLALA
                     NNSTLTIGTIDEIQKLHIRTVPLYESPRKICYQEVSQCFGVLSSRIEVQDTSGGTTALR
                     PSASTQALSSSVSSSKLFSSSTAPHETSFGEEVEVHNLLIIDQHTFEVLHAHQFLQNEY
                     ALSLVSCKLGKDPNTYFIVGTAMVYPEEAEPKQGRIVVFQYSDGKLQTVAEKEVKGAVY
                     SMVEFNGKLLASINSTVRLYEWTTEKELRTECNHYNNIMALYLKTKGDFILVGDLMRSV
                     LLLAYKPMEGNFEEIARDFNPNWMSAVEILDDDNFLGAENAFNLFVCQKDSAATTDEER
                     QHLQEVGLFHLGEFVNVFCHGSLVMQNLGETSTPTQGSVLFGTVNGMIGLVTSLSESWY
                     NLLLDMQNRLNKVIKSVGKIEHSFWRSFHTERKTEPATGFIDGDLIESFLDISRPKMQE
                     VVANLQYDDGSGMKREATADDLIKVVEELTRIH"
     promoter        complement(4373..4391)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4411..4428)
                     /label=BGH-R
     polyA_signal    4417..4641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4432..4450)
                     /label=pcDNA3.1-RP_1
     primer_bind     complement(4479..4499)
                     /label=BGH-seqR2
     rep_origin      4687..5115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5129..5458
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5309..5444
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5525..6319
                     /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    6493..6614
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6663..6679)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6687..6703
                     /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(6711..6741)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6756..6777
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7065..7653)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7824..8684)
                     /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"
     promoter        complement(8685..8789)
                     /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 ggagacatgg actacaaaga
      901 cgatgacgat aaaccaagct tggtaccggg tatgtcgtac aactacgtgg taacggccca
      961 gaagcccacc gccgtgaacg gctgcgtgac cggacacttt acttcggccg aagacttaaa
     1021 cctgttgatt gccaaaaaca cgagattaga gatctatgtg gtcaccgccg aggggcttcg
     1081 gcccgtcaaa gaggtgggca tgtatgggaa gattgcggtc atggagcttt tcaggcccaa
     1141 gggggagagc aaggacctgc tgtttatctt gacagcgaag tacaatgcct gcatcctgga
     1201 gtataaacag agtggcgaga gcattgacat cattacgcga gcccatggca atgtccagga
     1261 ccgcattggc cgcccctcag agaccggcat tattggcatc attgaccctg agtgccggat
     1321 gattggcctg cgtctctatg atggcctttt caaggttatt ccactagatc gcgataataa
     1381 agaactcaag gccttcaaca tccgcctgga ggagctgcat gtcattgatg tcaagttcct
     1441 atatggttgc caagcaccta ctatttgctt tgtctaccag gaccctcagg ggcggcacgt
     1501 aaaaacctat gaggtgtctc tccgagaaaa ggaattcaat aagggccctt ggaaacagga
     1561 aaatgtcgaa gctgaagctt ccatggtgat cgcagtccca gagccctttg ggggggccat
     1621 catcattgga caggagtcaa tcacctatca caatggtgac aaatacctgg ctattgcccc
     1681 tcctatcatc aagcaaagca cgattgtgtg ccacaatcga gtggacccta atggctcaag
     1741 atacctgctg ggagacatgg aaggccggct cttcatgctg cttttggaga aggaggaaca
     1801 gatggatggc accgtcactc tcaaggatct ccgtgtagaa ctccttggag agacctctat
     1861 tgctgagtgc ttgacatacc ttgataatgg tgttgtgttt gtcgggtctc gcctgggtga
     1921 ctcccagctt gtgaagctca acgttgacag taatgaacaa ggctcctatg tagtggccat
     1981 ggaaaccttt accaacttag gacccattgt cgatatgtgc gtggtggacc tggagaggca
     2041 ggggcagggg cagctggtca cttgctctgg ggctttcaag gaaggttctt tgcggatcat
     2101 ccggaatgga attggaatcc acgagcatgc cagcattgac ttaccaggca tcaaaggatt
     2161 atggccactg cggtctgacc ctaatcgtga gactgatgac actttggtgc tctcttttgt
     2221 gggccagaca agagttctca tgttaaatgg agaggaggta gaagaaaccg aactgatggg
     2281 tttcgtggat gatcagcaga ctttcttctg tggcaacgtg gctcatcagc agcttatcca
     2341 gatcacttca gcatcggtga ggttggtctc tcaagaaccc aaagctctgg tcagtgaatg
     2401 gaaggagcct caggccaaga acatcagtgt ggcctcctgc aatagcagcc aggtggtggt
     2461 ggctgtaggc agggccctct actatctgca gatccatcct caggagctcc ggcagatcag
     2521 ccacacagag atggaacatg aagtggcttg cttggacatc accccattag gagacagcaa
     2581 tggactgtcc cctctttgtg ccattggcct ctggacggac atctcggctc gtatcttgaa
     2641 gttgccctct tttgaactac tgcacaagga gatgctgggt ggagagatca ttcctcgctc
     2701 catcctgatg accacctttg agagtagcca ttacctcctt tgtgccttgg gagatggagc
     2761 gcttttctac tttgggctca acattgagac aggtctgttg agcgaccgta agaaggtgac
     2821 tttgggcacc cagcccaccg tattgaggac ttttcgttct ctttctacca ccaacgtctt
     2881 tgcttgttct gaccgcccca ctgtcatcta tagcagcaac cacaaattgg tcttctcaaa
     2941 tgtcaacctc aaggaagtga actacatgtg tcccctcaat tcagatggct atcctgacag
     3001 cctggcgctg gccaacaata gcaccctcac cattggcacc atcgatgaga tccagaagct
     3061 gcacattcgc acagttcccc tctatgagtc tccaaggaag atctgctacc aggaagtgtc
     3121 ccagtgtttc ggggtcctct ccagccgcat tgaagtccaa gacacgagtg ggggcacgac
     3181 agccttgagg cccagcgcta gcacccaggc tctgtccagc agtgtaagct ccagcaagct
     3241 gttctccagc agcactgctc ctcatgagac ctcctttgga gaagaggtgg aggtgcacaa
     3301 cctacttatc attgaccaac acacctttga agtgcttcat gcccaccagt ttctgcagaa
     3361 tgaatatgcc ctcagtctgg tttcctgcaa gctgggcaaa gaccccaaca cttacttcat
     3421 tgtgggcaca gcaatggtgt atcctgaaga ggcagagccc aagcagggtc gcattgtggt
     3481 ctttcagtat tcggatggaa aactacagac tgtggctgaa aaggaagtga aaggggccgt
     3541 gtactctatg gtggaattta acgggaagct gttagccagc atcaatagca cggtgcggct
     3601 ctatgagtgg acaacagaga aggagctgcg cactgagtgc aaccactaca acaacatcat
     3661 ggccctctac ctgaagacca agggcgactt catcctggtg ggcgacctta tgcgctcagt
     3721 gctgctgctt gcctacaagc ccatggaagg aaactttgaa gagattgctc gagactttaa
     3781 tcccaactgg atgagtgctg tggaaatctt ggatgatgac aattttctgg gggctgaaaa
     3841 tgcctttaac ttgtttgtgt gtcaaaagga tagcgctgcc accactgacg aggagcggca
     3901 gcacctccag gaggttggtc ttttccacct gggcgagttt gtcaatgtct tttgccacgg
     3961 ctctctggta atgcagaatc tgggtgagac ttccaccccc acacaaggct cggtgctctt
     4021 cggcacggtc aacggcatga tagggctggt gacctcactg tcagagagct ggtacaacct
     4081 cctgctggac atgcagaatc gactcaataa agtcatcaaa agtgtgggga agatcgagca
     4141 ctccttctgg agatcctttc acaccgagcg gaagacagaa ccagccacag gtttcatcga
     4201 cggtgacttg attgagagtt tcctggatat tagccgcccc aagatgcagg aggtggtggc
     4261 aaacctacag tatgacgatg gcagcggtat gaagcgagag gccactgcag acgacctcat
     4321 caaggttgtg gaggagctaa ctcggataca ttagatatct agagggccct attctatagt
     4381 gtcacctaaa tgctagagct cgctgatcag cctcgactgt gccttctagt tgccagccat
     4441 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     4501 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     4561 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4621 gggatgcggt gggctctatg gcttctgagg cggaaagaac cagctggggc tctagggggt
     4681 atccccacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     4741 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     4801 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     4861 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     4921 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     4981 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     5041 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     5101 aatttaacgc gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5161 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg
     5221 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     5281 aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca
     5341 ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc
     5401 ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa
     5461 gctcccggga gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt
     5521 tcgcatgatt gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct
     5581 attcggctat gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct
     5641 gtcagcgcag gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga
     5701 actgcaggac gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc
     5761 tgtgctcgac gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg
     5821 gcaggatctc ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc
     5881 aatgcggcgg ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca
     5941 tcgcatcgag cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga
     6001 cgaagagcat caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc
     6061 cgacggcgag gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga
     6121 aaatggccgc ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca
     6181 ggacatagcg ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg
     6241 cttcctcgtg ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct
     6301 tcttgacgag ttcttctgag cgggactctg gggttcgaaa tgaccgacca agcgacgccc
     6361 aacctgccat cacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     6421 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     6481 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     6541 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     6601 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     6661 tggtcatagc tgtttcctgt gtgaaattgt tatccgctca caattccaca caacatacga
     6721 gccggaagca taaagtgtaa agcctggggt gcctaatgag tgagctaact cacattaatt
     6781 gcgttgcgct cactgcccgc tttccagtcg ggaaacctgt cgtgccagct gcattaatga
     6841 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     6901 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     6961 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     7021 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     7081 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     7141 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     7201 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     7261 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     7321 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     7381 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     7441 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     7501 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     7561 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     7621 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     7681 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     7741 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     7801 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     7861 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     7921 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     7981 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     8041 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     8101 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     8161 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     8221 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     8281 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     8341 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     8401 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     8461 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     8521 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     8581 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     8641 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     8701 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     8761 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc
//
