LOCUS       Exported                5834 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 5834)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, May 26, 2020 from SnapGene 5.1.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5834
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     promoter        298..501
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate earlypromoter"
                     /note="color: #ffffff; direction: RIGHT"
     promoter        546..564
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: RIGHT"
     misc_feature    597
                     /label=Apa1
                     /note="color: #f3ff4a"
     CDS             640..1305
                     /codon_start=1
                     /label=coding
                     /note="color: #4cff46"
                     /translation="MGLFGKTQEKPPKELVNEWSLKIRKEMRVVDRQIRDIQREEEKVK
                     RSVKDAAKKGQKDVCVVLAKEMIRSRKAVSKLYASKAHMNSVLMGMKNQLAVLRVAGSL
                     QKSTEVMKAMQSLVKIPEIQATMRELSKEMMKAGIIEEMLEDTFESMDDQEEMEEEAEM
                     EIDKILFEITAGALGKAPSKVTDALPEPEPSGAMAASEDEEEEEEALEAMQSRLATLRS
                     "
     misc_feature    880..912
                     /label=Primers 225 and 226
                     /note="color: #62ff17"
     misc_feature    1286..1305
                     /label=Reverse PRimer
                     /note="color: #ff98f9"
     misc_feature    1306..1332
                     /label=HA tag
                     /note="color: #00ffff"
     misc_feature    1333..1335
                     /label=STOP
                     /note="color: #ff2326"
     misc_feature    1336
                     /label=HindIII
                     /note="color: #fbff65"
     polyA_signal    1431..1655
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     rep_origin      1701..2129
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of 
                     replication;arrowindicates direction of (+) strand 
                     synthesis"
                     /note="color: #ffff00"
     promoter        2143..2472
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      2323..2458
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     CDS             2539..3333
                     /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    3507..3628
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(3677..3693)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    3701..3717
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operatortoinhibit
                     transcription in E. coli. This inhibition can be relieved 
                     by adding lactose or isopropyl-beta-D-thiogalactopyranoside
                     (IPTG)."
                     /note="color: #31849b"
     promoter        complement(3725..3755)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 3725 .. 3731 / #ffffff / -10
                        2: 3732 .. 3749 / #ffffff
                        3: 3750 .. 3755 / #ffffff / -35"
     protein_bind    3770..3791
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in thepresenceof
                     cAMP."
                     /note="color: #31849b"
     rep_origin      complement(4079..4667)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(4838..5698)
                     /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: 4838 .. 5629 / #ccffcc
                        2: 5630 .. 5698 / #ccffcc / signal sequence"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(5699..5803)
                     /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 cgtcatgacc
      241 ttatgggact ttcctacttg gcagtacatc tacgtattag tcatcgctat taccatggtg
      301 atgcggtttt ggcagtacat caatgggcgt ggatagcggt ttgactcacg gggatttcca
      361 agtctccacc ccattgacgt caatgggagt ttgttttggc accaaaatca acgggacttt
      421 ccaaaatgtc gtaacaactc cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg
      481 gaggtctata taagcagagc tctctggcta actagagaac ccactgctta ctggcttatc
      541 gaaattaata cgactcacta tagggagacc caagctggct agcgtttaaa cgggcccggg
      601 aacggaaagc cggaaggagc aagacgagtt cagttcgcca tggggctgtt tggaaaaacc
      661 caggagaagc cacccaaaga actggtcaat gaatggtcat tgaagataag aaaggaaatg
      721 agagttgttg acaggcaaat aagggatatc caaagagaag aagaaaaagt gaaacgatct
      781 gtgaaagatg cagccaagaa gggccagaag gatgtctgtg tagttctggc caaggagatg
      841 atcaggtcaa ggaaggccgt gagcaagcta tatgcatcca aagcgcacat gaattcggtg
      901 ctcatgggga tgaagaacca gctcgcggtc ttgcgagtgg ctggttccct gcagaagagc
      961 acagaagtga tgaaggccat gcaaagtctt gtgaagattc cagaaattca ggccaccatg
     1021 agggagctat ccaaagaaat gatgaaggct gggatcatag aggagatgtt agaggacact
     1081 tttgaaagca tggacgatca ggaagaaatg gaggaagaag cagaaatgga aattgacaaa
     1141 attctatttg aaattacagc aggggccttg ggcaaagcac ccagtaaagt gactgatgcc
     1201 cttccagagc cagaaccttc gggagcgatg gctgcctcag aggatgagga ggaggaggaa
     1261 gaagctctgg aggccatgca gtcccggctg gccacactcc gaagctaccc atacgatgtt
     1321 ccagattacg cttaaaagct tgctagcaga tctttttccc tctgccaaaa attatgggga
     1381 catcatgaag ccccttgagc acttaagttt aaaccgctga tcagcctcga ctgtgccttc
     1441 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1501 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1561 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1621 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     1681 gggctctagg gggtatcccc acgcgccctg tagcggcgca ttaagcgcgg cgggtgtggt
     1741 ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc ctttcgcttt
     1801 cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa atcgggggct
     1861 ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac ttgattaggg
     1921 tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt tgacgttgga
     1981 gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca accctatctc
     2041 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     2101 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     2161 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     2221 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     2281 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2341 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2401 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2461 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcaagagaca
     2521 ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc tccggccgct
     2581 tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg ctctgatgcc
     2641 gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac cgacctgtcc
     2701 ggtgccctga atgaactgca ggacgaggca gcgcggctat cgtggctggc cacgacgggc
     2761 gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg gctgctattg
     2821 ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga gaaagtatcc
     2881 atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg cccattcgac
     2941 caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg tcttgtcgat
     3001 caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt cgccaggctc
     3061 aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc ctgcttgccg
     3121 aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg gctgggtgtg
     3181 gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga gcttggcggc
     3241 gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc gcagcgcatc
     3301 gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc gaaatgaccg
     3361 accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc ttctatgaaa
     3421 ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag cgcggggatc
     3481 tcatgctgga gttcttcgcc caccccaact tgtttattgc agcttataat ggttacaaat
     3541 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     3601 gtttgtccaa actcatcaat gtatcttatc atgtctgtat accgtcgacc tctagctaga
     3661 gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc
     3721 cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct
     3781 aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc
     3841 agctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt gggcgctctt
     3901 ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag
     3961 ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca
     4021 tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt
     4081 tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc
     4141 gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct
     4201 ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg
     4261 tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca
     4321 agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact
     4381 atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta
     4441 acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta
     4501 actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct
     4561 tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt
     4621 tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga
     4681 tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca
     4741 tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat
     4801 caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg
     4861 cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt
     4921 agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag
     4981 acccacgctc accggctcca gatttatcag caataaacca gccagccgga agggccgagc
     5041 gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag
     5101 ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca
     5161 tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa
     5221 ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga
     5281 tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata
     5341 attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca
     5401 agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg
     5461 ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg
     5521 ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg
     5581 cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag
     5641 gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac
     5701 tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca
     5761 tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag
     5821 tgccacctga cgtc
//
