LOCUS       Exported                5893 bp ds-DNA     circular SYN 13-JUL-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5893)
  AUTHORS   Yaxin Li
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jul 13, 2020 from SnapGene Viewer 5.1.4.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5893
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     rep_origin      46..474
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        488..817
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      668..803
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             884..1678
                     /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    1852..1973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2022..2038)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2046..2062
                     /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(2070..2100)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2115..2136
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2424..3009)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3180..4040)
                     /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(4041..4145)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4411..4790
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4791..4994
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        5039..5057
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5099..5599
                     /codon_start=1
                     /product="8xHis affinity tag"
                     /label=insert
                     /translation="MKNHLLFWGVLAVFIKAVHVKAQEDERIVLVDNKCKCARITSRII
                     RSSEDPNEDIVERNIRIIVPLNNRENISDPTSPLRTRFVYHLSDLCKKCDPTEVELDNQ
                     IVTATQSNICDEDSATETCYTYDRNKCYTAVVPLVYGGETKMVETALTPDACYPDHHHH
                     HHHH"
     CDS             5612..5635
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     polyA_signal    5669..5893
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
ORIGIN
        1 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
       61 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
      121 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
      181 cccgtcaagc tctaaatcgg gggctccctt tagggttccg atttagtgct ttacggcacc
      241 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
      301 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
      361 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgccga
      421 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
      481 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
      541 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
      601 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
      661 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
      721 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
      781 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
      841 cattttcgga tctgatcaag agacaggatg aggatcgttt cgcatgattg aacaagatgg
      901 attgcacgca ggttctccgg ccgcttgggt ggagaggcta ttcggctatg actgggcaca
      961 acagacaatc ggctgctctg atgccgccgt gttccggctg tcagcgcagg ggcgcccggt
     1021 tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa ctgcaggacg aggcagcgcg
     1081 gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga
     1141 agcgggaagg gactggctgc tattgggcga agtgccgggg caggatctcc tgtcatctca
     1201 ccttgctcct gccgagaaag tatccatcat ggctgatgca atgcggcggc tgcatacgct
     1261 tgatccggct acctgcccat tcgaccacca agcgaaacat cgcatcgagc gagcacgtac
     1321 tcggatggaa gccggtcttg tcgatcagga tgatctggac gaagagcatc aggggctcgc
     1381 gccagccgaa ctgttcgcca ggctcaaggc gcgcatgccc gacggcgagg atctcgtcgt
     1441 gacccatggc gatgcctgct tgccgaatat catggtggaa aatggccgct tttctggatt
     1501 catcgactgt ggccggctgg gtgtggcgga ccgctatcag gacatagcgt tggctacccg
     1561 tgatattgct gaagagcttg gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat
     1621 cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc
     1681 gggactctgg ggttcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc
     1741 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     1801 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     1861 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     1921 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     1981 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg
     2041 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     2101 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     2161 ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     2221 ggcggtttgc gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     2281 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     2341 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     2401 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     2461 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     2521 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     2581 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     2641 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     2701 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     2761 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     2821 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     2881 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     2941 caaaccaccg ctggtagcgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3001 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3061 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3121 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3181 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3241 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3301 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3361 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3421 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3481 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3541 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3601 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3661 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3721 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3781 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3841 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3901 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3961 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4021 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4081 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4141 ccgcgcacat ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc
     4201 ccctatggtg cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc
     4261 tccctgcttg tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa
     4321 ggcaaggctt gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc
     4381 ttcgcgatgt acgggccaga tatacgcgtt gacattgatt attgactagt tattaatagt
     4441 aatcaattac ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta
     4501 cggtaaatgg cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga
     4561 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
     4621 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta
     4681 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg
     4741 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
     4801 tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
     4861 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
     4921 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
     4981 atataagcag agctctctgg ctaactagag aacccactgc ttactggctt atcgaaatta
     5041 atacgactca ctatagggag acccaagctg gctagcgttt aaacttaagc ttgccaccat
     5101 gaagaaccat ttgcttttct ggggagtcct ggcggttttt attaaggctg ttcatgtgaa
     5161 agcccaagaa gatgaaagga ttgttcttgt tgacaacaaa tgtaagtgtg cccggattac
     5221 ttccaggatc atccgttctt ccgaagatcc taatgaggac attgtggaga gaaacatccg
     5281 aattattgtt cctctgaaca acagggagaa tatctctgat cccacctcac cattgagaac
     5341 cagatttgtg taccatttgt ctgacctctg taaaaaatgt gatcctacag aagtggagct
     5401 ggataatcag atagttactg ctacccagag caatatctgt gatgaagaca gtgctacaga
     5461 gacctgctac acttatgaca gaaacaagtg ctacacagct gtggtcccac tcgtatatgg
     5521 tggtgagacc aaaatggtgg aaacagcctt aaccccagat gcctgctatc ctgaccacca
     5581 tcaccatcac catcaccact aatctagagt cgactacaag gacgacgatg acaagtaggg
     5641 cccgtttaaa cccgctgatc agcctcgact gtgccttcta gttgccagcc atctgttgtt
     5701 tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca ctcccactgt cctttcctaa
     5761 taaaatgagg aaattgcatc gcattgtctg agtaggtgtc attctattct ggggggtggg
     5821 gtggggcagg acagcaaggg ggaggattgg gaagacaata gcaggcatgc tggggatgcg
     5881 gtgggctcta tgg
//
