LOCUS       Exported                6564 bp ds-DNA     circular SYN 12-NOV-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6564)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 30, 2020 from SnapGene Viewer 5.2.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6564
                     /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             898..945
                     /codon_start=1
                     /product="11th beta-strand of superfolder GFP"
                     /label=GFP11
                     /note="associates with GFP(1-10), consisting of the first 
                     10 beta-strands, to reconstitute fluorescent GFP (Cabantous
                     et al., 2005; Kamiyama et al., 2016)"
                     /translation="RDHMVLHEYVNAAGIT"
     CDS             1126..1182
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     promoter        complement(2116..2134)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    2160..2384
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2430..2858
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2872..3202
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3053..3188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3269..4063
                     /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    4237..4358
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4407..4423)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4431..4447
                     /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(4455..4485)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4500..4521
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4809..5397)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5568..6428)
                     /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(6429..6533)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 atgggtggac 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 gcttatgcgg
      901 gaccacatgg tgctgcacga gtacgtgaac gccgccggca tcaccggagg cgatggaggc
      961 agcggaggag gctccaagct gtggtggccc ttcgagatgt ctcctggcgt gctgacattc
     1021 gccatcatct ggccctttat cgcccagtgg ctggtgtatc tgtactatca gcggagaagg
     1081 cgcaagctga atggatccct gaaggagctg gcatgcggca gcggagcaac caacttctcc
     1141 ctgctgaagc aggcaggcga cgtggaggag aatcccggcc ctcggtctga attcatgagc
     1201 aagggagagg agctgttcac cggagtggtg cctatcctgg tggagctgga cggcgatgtg
     1261 aatggccaca agttttccgt gcgcggagag ggagagggcg acgcaacaat cggcaagctg
     1321 accctgaagt tcatctgcac cacaggcaag ctgcccgtgc cttggccaac cctggtgacc
     1381 acactgggct acggcctgca gtgttttgcc cggtatccag accacatgaa gagacacgat
     1441 ttctttaagt ccgccatgcc cgagggctac gtgcaggaga ggacaatctc tttcaaggac
     1501 gatggcaagt ataagacacg cgccgtggtg aagtttgagg gcgacaccct ggtgaaccgg
     1561 atcgagctga agggcacaga cttcaaggag gatggcaata tcctgggcca caagctggag
     1621 tacaacttta attcccacaa cgtgtatatc accgccgata agcagaagaa cggcatcaag
     1681 gccaatttca cagtgcggca caatgtggag gacggctctg tgcagctggc cgatcactac
     1741 cagcagaaca cccctatcgg cgacggccct gtgctgctgc ctgataatca ctacctgtct
     1801 tatcagaccg tgctgagcaa ggacccaaac gagaagggca caggaggcga tggaggcagc
     1861 ggaggaggct ccaagctgag agtgttcctg gccctgccca tcatcatggt ggtggccttt
     1921 tctatgtgca tcatctgtct gctgatggca ggcgacacct ggacagagac cctggcctac
     1981 gtgctgtttt ggggcgtggc cagcatcggc acattcttta tcatcctgta taacggcaag
     2041 gacttcgtgg atgcccccag gctggtgcag aaggagaaga tcgattgact cgagcatgca
     2101 tctagagggc cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac
     2161 tgtgccttct agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct
     2221 ggaaggtgcc actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct
     2281 gagtaggtgt cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg
     2341 ggaagacaat agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag
     2401 aaccagctgg ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc
     2461 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     2521 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     2581 tcggggcatc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     2641 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     2701 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     2761 ccctatctcg gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt
     2821 aaaaaatgag ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag
     2881 ttagggtgtg gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct
     2941 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3001 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     3061 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     3121 tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt
     3181 tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga
     3241 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3301 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3361 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3421 cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc
     3481 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3541 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3601 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3661 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3721 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3781 cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3841 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3901 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3961 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     4021 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     4081 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     4141 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4201 cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat
     4261 ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat
     4321 tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc
     4381 tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     4441 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     4501 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     4561 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     4621 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     4681 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     4741 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     4801 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4861 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4921 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4981 tcgggaagcg tggcgctttc tcaatgctca cgctgtaggt atctcagttc ggtgtaggtc
     5041 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5101 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5161 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5221 tggtggccta actacggcta cactagaagg acagtatttg gtatctgcgc tctgctgaag
     5281 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5341 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5401 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     5461 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     5521 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     5581 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     5641 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     5701 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     5761 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     5821 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     5881 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     5941 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6001 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6061 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6121 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6181 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6241 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6301 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6361 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6421 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     6481 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     6541 ccccgaaaag tgccacctga cgtc
//
