LOCUS       Exported                8182 bp DNA     circular SYN 19-MAY-2020
DEFINITION  -.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8182)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 8, 2016 from SnapGene 2.3.2
            http://www.snapgene.com
REFERENCE   2  (bases 1 to 8182)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 9, 2022 from SnapGene 5.3.0
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Exported 
            Nov 8, 2016 from SnapGene 2.3.2
            http://www.snapgene.com"
FEATURES             Location/Qualifiers
     source          1..8182
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        237..616
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     enhancer        237..616
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        617..820
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        618..816
                     /note="CMV promoter"
                     /note="human cytomegalovirus (CMV) immediate early  
                     promoter"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     LTR             834..1014
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="HIV-1 Psi"
                     /note="packaging signal of human immunodeficiency virus  
                     type 1"
     misc_feature    1061..1186
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus 
                     type 1"
     misc_feature    1685..1918
                     /label=RRE
                     /note="RRE"
                     /note="The Rev response element (RRE) of HIV-1 allows for  
                     Rev-dependent mRNA export from the nucleus to the  
                     cytoplasm."
     misc_feature    1685..1918
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    2445..2562
                     /label=cPPT/CTS
                     /note="cPPT/CTS"
                     /note="central polypurine tract and central termination  
                     sequence of HIV-1"
     misc_feature    2445..2562
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination 
                     sequence of HIV-1"
     primer_bind     2625..2654
                     /label=pAF
     promoter        2667..2878
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     primer_bind     2887..2916
                     /label=gAF
     primer_bind     complement(2887..2916)
                     /label=pAR
     regulatory      2917..2926
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2923..3639
                     /codon_start=1
                     /label=GFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     misc_feature    3643..3672
                     /label=pBF
                     /note="pBF"
     misc_feature    3643..3672
                     /label=gAR
                     /note="gAR"
     primer_bind     3650..3691
                     /label=pBF-PGK-F
     promoter        3673..4177
                     /label=PGK
                     /note="PGK"
     primer_bind     3673..3693
                     /label=FSR-16 (w/o Gibson Adapters)
                     /note="FSR-16 (w/o Gibson Adapters)"
     misc_feature    4100..4120
                     /label=SANGER-END
     primer_bind     complement(4158..4207)
                     /label=Primer 2
     primer_bind     4178..4235
                     /label=gBF-Neo
     primer_bind     complement(4178..4207)
                     /label=pBR
     primer_bind     4178..4207
                     /label=gBF
     misc_feature    4208..4213
                     /label=KOZAK
                     /note="KOZAK"
     CDS             4214..5008
                     /codon_start=1
                     /label=Neo
                     /note="/vntifkey=""4"""
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    4954..4991
                     /label=SANGER-START
     primer_bind     complement(4987..5048)
                     /label=gBR-Neo-R
     primer_bind     complement(5019..5048)
                     /label=gBR
     misc_feature    5071..5659
                     /label=WPRE
                     /note="WPRE"
                     /note="woodchuck hepatitis virus posttranscriptional  
                     regulatory element"
     misc_feature    5071..5659
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     CDS             complement(5542..5553)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(5662..5678)
                     /label=KS primer
                     /note="KS primer"
                     /note="common sequencing primer, one of multiple similar  
                     variants"
     primer_bind     complement(5662..5678)
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             6188..6368
                     /label=5' LTR (truncated)
                     /note="5' LTR (truncated)"
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     LTR             6188..6368
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     rep_origin      6430..7018
                     /label=ori
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of  
                     replication"
     rep_origin      complement(6430..7018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7189..8049)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=bla
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     CDS             complement(7189..8049)
                     /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(8050..8154)
                     /gene="bla"
                     /label=bla promoter
                     /note="AmpR promoter"
     promoter        complement(8050..8154)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
       61 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
      121 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
      181 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca
      241 ttgattattg actagttatt aatagtaatc aattacgggg tcattagttc atagcccata
      301 tatggagttc cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga
      361 cccccgccca ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt
      421 ccattgacgt caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt
      481 gtatcatatg ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca
      541 ttatgcccag tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt
      601 catcgctatt accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt
      661 tgactcacgg ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca
      721 ccaaaatcaa cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg
      781 cggtaggcgt gtacggtggg aggtctatat aagcagcgcg ttttgcctgt actgggtctc
      841 tctggttaga ccagatctga gcctgggagc tctctggcta actagggaac ccactgctta
      901 agcctcaata aagcttgcct tgagtgcttc aagtagtgtg tgcccgtctg ttgtgtgact
      961 ctggtaacta gagatccctc agaccctttt agtcagtgtg gaaaatctct agcagtggcg
     1021 cccgaacagg gacttgaaag cgaaagggaa accagaggag ctctctcgac gcaggactcg
     1081 gcttgctgaa gcgcgcacgg caagaggcga ggggcggcga ctggtgagta cgccaaaaat
     1141 tttgactagc ggaggctaga aggagagaga tgggtgcgag agcgtcagta ttaagcgggg
     1201 gagaattaga tcgcgatggg aaaaaattcg gttaaggcca gggggaaaga aaaaatataa
     1261 attaaaacat atagtatggg caagcaggga gctagaacga ttcgcagtta atcctggcct
     1321 gttagaaaca tcagaaggct gtagacaaat actgggacag ctacaaccat cccttcagac
     1381 aggatcagaa gaacttagat cattatataa tacagtagca accctctatt gtgtgcatca
     1441 aaggatagag ataaaagaca ccaaggaagc tttagacaag atagaggaag agcaaaacaa
     1501 aagtaagacc accgcacagc aagcggccgg ccgcgctgat cttcagacct ggaggaggag
     1561 atatgaggga caattggaga agtgaattat ataaatataa agtagtaaaa attgaaccat
     1621 taggagtagc acccaccaag gcaaagagaa gagtggtgca gagagaaaaa agagcagtgg
     1681 gaataggagc tttgttcctt gggttcttgg gagcagcagg aagcactatg ggcgcagcgt
     1741 caatgacgct gacggtacag gccagacaat tattgtctgg tatagtgcag cagcagaaca
     1801 atttgctgag ggctattgag gcgcaacagc atctgttgca actcacagtc tggggcatca
     1861 agcagctcca ggcaagaatc ctggctgtgg aaagatacct aaaggatcaa cagctcctgg
     1921 ggatttgggg ttgctctgga aaactcattt gcaccactgc tgtgccttgg aatgctagtt
     1981 ggagtaataa atctctggaa cagatttgga atcacacgac ctggatggag tgggacagag
     2041 aaattaacaa ttacacaagc ttaatacact ccttaattga agaatcgcaa aaccagcaag
     2101 aaaagaatga acaagaatta ttggaattag ataaatgggc aagtttgtgg aattggttta
     2161 acataacaaa ttggctgtgg tatataaaat tattcataat gatagtagga ggcttggtag
     2221 gtttaagaat agtttttgct gtactttcta tagtgaatag agttaggcag ggatattcac
     2281 cattatcgtt tcagacccac ctcccaaccc cgaggggacc cgacaggccc gaaggaatag
     2341 aagaagaagg tggagagaga gacagagaca gatccattcg attagtgaac ggatcggcac
     2401 tgcgtgcgcc aattctgcag acaaatggca gtattcatcc acaattttaa aagaaaaggg
     2461 gggattgggg ggtacagtgc aggggaaaga atagtagaca taatagcaac agacatacaa
     2521 actaaagaat tacaaaaaca aattacaaaa attcaaaatt ttcgggttta ttacagggac
     2581 agcagagatc cagtttggtt agtaccgggc ccgggtcgac tagtgatcag tgtgagggag
     2641 tgtaaagctg gtttaaaccc tgcagggggc agagcgcaca tcgcccacag tccccgagaa
     2701 gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg gggtaaactg
     2761 ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag aaccgtatat
     2821 aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca gaacacagcg
     2881 ccggtgctaa ctcgaacgct agctgtgcga tcgtttgcca ccatggtgag caagggcgag
     2941 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
     3001 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
     3061 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
     3121 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
     3181 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
     3241 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     3301 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     3361 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     3421 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     3481 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     3541 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     3601 gccgccggga tcactctcgg catggacgag ctgtacaagt aaaaaccgct gttcctagga
     3661 atcccgaggc ctctaccggg taggggaggc gcttttccca aggcagtctg gagcatgcgc
     3721 tttagcagcc ccgctgggca cttggcgcta cacaagtggc ctctggcctc gcacacattc
     3781 cacatccacc ggtaggcgcc aaccggctcc gttctttggt ggccccttcg cgccaccttc
     3841 tactcctccc ctagtcagga agttcccccc cgccccgcag ctcgcgtcgt gcaggacgtg
     3901 acaaatggaa gtagcacgtc tcactagtct cgtgcagatg gacagcaccg ctgagcaatg
     3961 gaagcgggta ggcctttggg gcagcggcca atagcagctt tgctccttcg ctttctgggc
     4021 tcagaggctg ggaaggggtg ggtccggggg cgggctcagg ggcgggctca ggggcggggc
     4081 gggcgcccga aggtcctccg gaggcccggc attctgcacg cttcaaaagc gcacgtctgc
     4141 cgcgctgttc tcctcttcct catctccggg cctttcggac ccgacattag cgctacagct
     4201 taagcgggcc accatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt
     4261 ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt
     4321 gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc
     4381 cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc
     4441 ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga
     4501 agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat
     4561 ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca
     4621 agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga
     4681 tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc
     4741 gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat
     4801 catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga
     4861 ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg
     4921 ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt
     4981 ctatcgcctt cttgacgagt tcttctgagg atccgtttaa acgttgttgt ttggggttga
     5041 attactcttc tagaccgcgg cgcgcccgat aatcaacctc tggattacaa aatttgtgaa
     5101 agattgactg gtattcttaa ctatgttgct ccttttacgc tatgtggata cgctgcttta
     5161 atgcctttgt atcatgctat tgcttcccgt atggctttca ttttctcctc cttgtataaa
     5221 tcctggttgc tgtctcttta tgaggagttg tggcccgttg tcaggcaacg tggcgtggtg
     5281 tgcactgtgt ttgctgacgc aacccccact ggttggggca ttgccaccac ctgtcagctc
     5341 ctttccggga ctttcgcttt ccccctccct attgccacgg cggaactcat cgccgcctgc
     5401 cttgcccgct gctggacagg ggctcggctg ttgggcactg acaattccgt ggtgttgtcg
     5461 gggaaatcat cgtcctttcc ttggctgctc gcctgtgttg ccacctggat tctgcgcggg
     5521 acgtccttct gctacgtccc ttcggccctc aatccagcgg accttccttc ccgcggcctg
     5581 ctgccggctc tgcggcctct tccgcgtctt cgccttcgcc ctcagacgag tcggatctcc
     5641 ctttgggccg cctccccgca tcgataccgt cgacctcgat cgagacctag aaaaacatgg
     5701 agcaatcaca agtagcaata cagcagctac caatgctgat tgtgcctggc tagaagcaca
     5761 agaggaggag gaggtgggtt ttccagtcac acctcaggta cctttaagac caatgactta
     5821 caaggcagct gtagatctta gccacttttt aaaagaaaag gggggactgg aagggctaat
     5881 tcactcccaa cgaagacaag atatccttga tctgtggatc taccacacac aaggctactt
     5941 ccctgattgg cagaactaca caccagggcc agggatcaga tatccactga cctttggatg
     6001 gtgctacaag ctagtaccag ttgagcaaga gaaggtagaa gaagccaatg aaggagagaa
     6061 cacccgcttg ttacaccctg tgagcctgca tgggatggat gacccggaga gagaagtatt
     6121 agagtggagg tttgacagcc gcctagcatt tcatcacatg gcccgagagc tgcatccgga
     6181 ctgtactggg tctctctggt tagaccagat ctgagcctgg gagctctctg gctaactagg
     6241 gaacccactg cttaagcctc aataaagctt gccttgagtg cttcaagtag tgtgtgcccg
     6301 tctgttgtgt gactctggta actagagatc cctcagaccc ttttagtcag tgtggaaaat
     6361 ctctagcagc atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     6421 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     6481 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     6541 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     6601 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     6661 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     6721 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     6781 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     6841 gtggtggcct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     6901 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     6961 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     7021 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg
     7081 gattttggtc atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg
     7141 aagttttaaa tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt
     7201 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
     7261 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
     7321 gataccgcga gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
     7381 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
     7441 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
     7501 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
     7561 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
     7621 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
     7681 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
     7741 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
     7801 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
     7861 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
     7921 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
     7981 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
     8041 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     8101 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     8161 tccccgaaaa gtgccacctg ac
//
