LOCUS       Exported                5734 bp ds-DNA     circular SYN 26-MAY-2020
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5734)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, May 26, 2020 from SnapGene 5.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5734
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     promoter        446..464
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    497
                     /label=Apa1
                     /note="color: #f3ff4a"
     CDS             540..1205
                     /codon_start=1
                     /label=coding
                     /note="color: #4cff46"
                     /translation="MGLFGKTQEKPPKELVNEWSLKIRKEMRVVDRQIRDIQREEEKVK
                     RSVKDAAKKGQKDVCVVLAKEMIRSRKAVSKLYASKAHMNSVLMGMKNQLAVLRVAGSL
                     QKSTEVMKAMQSLVKIPEIQATMRELSKEMMKAGIIEEMLEDTFESMDDQEEMEEEAEM
                     EIDKILFEITAGALGKAPSKVTDALPEPEPSGAMAASEDEEEEEEALEAMQSRLATLRS
                     "
     misc_feature    780..812
                     /label=Primers 225 and 226
                     /note="color: #62ff17"
     misc_feature    1186..1205
                     /label=Reverse PRimer
                     /note="color: #ff98f9"
     misc_feature    1206..1232
                     /label=HA tag
                     /note="color: #00ffff"
     misc_feature    1233..1235
                     /label=STOP
                     /note="color: #ff2326"
     polyA_signal    1331..1555
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      1601..2029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; 
                     arrowindicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
     promoter        2043..2372
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2223..2358
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             2439..3233
                     /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))"
                     /note="color: #ccffcc"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    3407..3528
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(3577..3593)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    3601..3617
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator 
                     toinhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(3625..3655)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 3625 .. 3631 / #ffffff / -10
                        2: 3632 .. 3649 / #ffffff
                        3: 3650 .. 3655 / #ffffff / -35"
     protein_bind    3670..3691
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the 
                     presenceof cAMP."
                     /note="color: #31849b"
     rep_origin      complement(3979..4567)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(4738..5598)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4738 .. 5529 / #ccffcc
                        2: 5530 .. 5598 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5599..5703)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
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 cgttagcggt
      241 ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt ttgttttggc
      301 accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg
      361 gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta actagagaac
      421 ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc caagctggct
      481 agcgtttaaa cgggcccggg aacggaaagc cggaaggagc aagacgagtt cagttcgcca
      541 tggggctgtt tggaaaaacc caggagaagc cacccaaaga actggtcaat gaatggtcat
      601 tgaagataag aaaggaaatg agagttgttg acaggcaaat aagggatatc caaagagaag
      661 aagaaaaagt gaaacgatct gtgaaagatg cagccaagaa gggccagaag gatgtctgtg
      721 tagttctggc caaggagatg atcaggtcaa ggaaggccgt gagcaagcta tatgcatcca
      781 aagcgcacat gaattcggtg ctcatgggga tgaagaacca gctcgcggtc ttgcgagtgg
      841 ctggttccct gcagaagagc acagaagtga tgaaggccat gcaaagtctt gtgaagattc
      901 cagaaattca ggccaccatg agggagctat ccaaagaaat gatgaaggct gggatcatag
      961 aggagatgtt agaggacact tttgaaagca tggacgatca ggaagaaatg gaggaagaag
     1021 cagaaatgga aattgacaaa attctatttg aaattacagc aggggccttg ggcaaagcac
     1081 ccagtaaagt gactgatgcc cttccagagc cagaaccttc gggagcgatg gctgcctcag
     1141 aggatgagga ggaggaggaa gaagctctgg aggccatgca gtcccggctg gccacactcc
     1201 gaagctaccc atacgatgtt ccagattacg cttaaaagct tgctagcaga tctttttccc
     1261 tctgccaaaa attatgggga catcatgaag ccccttgagc acttaagttt aaaccgctga
     1321 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     1381 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     1441 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     1501 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     1561 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     1621 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     1681 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     1741 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     1801 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     1861 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     1921 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     1981 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     2041 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     2101 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     2161 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2221 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2281 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     2341 gaggcttttt tggaggccta ggcttttgca aaaagctccc gggagcttgt atatccattt
     2401 tcggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     2461 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     2521 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     2581 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     2641 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     2701 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     2761 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     2821 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     2881 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     2941 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     3001 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     3061 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     3121 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     3181 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     3241 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     3301 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     3361 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     3421 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     3481 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     3541 accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa
     3601 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     3661 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     3721 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     3781 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     3841 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     3901 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     3961 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     4021 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     4081 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     4141 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     4201 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     4261 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     4321 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     4381 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     4441 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     4501 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     4561 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     4621 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     4681 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     4741 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     4801 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     4861 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     4921 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     4981 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     5041 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     5101 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     5161 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     5221 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     5281 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     5341 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     5401 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     5461 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     5521 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     5581 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     5641 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     5701 gcgcacattt ccccgaaaag tgccacctga cgtc
//
