LOCUS       Exported                6823 bp DNA     circular SYN 14-DEC-2021
DEFINITION  pcDNA3.1ZEO.seq, 5015 bases, 0 checksum.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6823)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2021 from SnapGene Viewer 6.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6823
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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             987..2690
                     /codon_start=1
                     /label=SEPT9_i3
                     /translation="MERDRISALKRSFEVEEVETPNSTPPRRVQTPLLRATVASSTQKF
                     QDLGVKNSEPSARHVDSLSQRSPKASLRRVELSGPKAAEPVSRRTELSIDISSKQVENA
                     GAIGPSRFGLKRAEVLGHKTPEPAPRRTEITIVKPQESAHRRMEPPASKVPEVPTAPAT
                     DAAPKRVEIQMPKPAEAPTAPSPAQTLENSEPAPVSQLQSRLEPKPQPPVAEATPRSQE
                     ATEAAPSCVGDMADTPRDAGLKQAPASRNEKAPVDFGYVGIDSILEQMRRKAMKQGFEF
                     NIMVVGQSGLGKSTLINTLFKSKISRKSVQPTSEERIPKTIEIKSITHDIEEKGVRMKL
                     TVIDTPGFGDHINNENCWQPIMKFINDQYEKYLQEEVNINRKKRIPDTRVHCCLYFIPA
                     TGHSLRPLDIEFMKRLSKVVNIVPVIAKADTLTLEERVHFKQRITADLLSNGIDVYPQK
                     EFDEDSEDRLVNEKFREMIPFAVVGSDHEYQVNGKRILGRKTKWGTIEVENTTHCEFAY
                     LRDLLIRTHMQNIKDITSSIHFEAYRVKRLNEGSSAMANGVEEKEPEAPEM"
     CDS             2691..2732
                     /codon_start=1
                     /label=14aa linker
                     /translation="IDGGGGSGGGGSSG"
     CDS             2733..2795
                     /codon_start=1
                     /label=GFP11
                     /translation="EKRDHMVLLEYVTAAGITDAS"
     polyA_signal    2836..3060
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3106..3534
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3548..3878
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3729..3864
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3926..3973
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3992..4366
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    4496..4617
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4533..4552)
                     /label=SV40pA-Rb
     primer_bind     complement(4533..4552)
                     /label=SV40pA-R
     primer_bind     complement(4666..4682)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4690..4706
                     /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(4714..4744)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4759..4780
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(5068..5656)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5109..5128)
                     /label=pTRETightBIREV
     primer_bind     complement(5661..5679)
                     /label=dual79i3CSseqREV
     CDS             complement(5827..6687)
                     /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"
     primer_bind     complement(6083..6106)
                     /label=pTAGsfGFPFOR
     primer_bind     6445..6462
                     /label=pTRETightBIFOR
     promoter        complement(6688..6792)
                     /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 gctggctagc
      901 gtttaaactt aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc
      961 agatatccag cacagtggcg gccgccatgg agagggaccg gatctcagcc ttgaaaagat
     1021 cttttgaggt cgaggaggtc gagacaccca actccacccc accccggagg gtccagactc
     1081 ccctactccg agccactgtg gccagctcca cccagaaatt ccaggacctg ggcgtgaaga
     1141 actcagaacc ctcggcccgc catgtggact ccctaagcca acgctccccc aaggcgtccc
     1201 tgcggagggt ggagctctcg ggccccaagg cggccgagcc ggtgtcccgg cgcactgagc
     1261 tgtccattga catctcgtcc aagcaggtgg agaacgccgg ggccatcggc ccgtcccggt
     1321 tcgggctcaa gagggccgag gtgttgggcc acaagacgcc agaaccggcc cctcggagga
     1381 cggagatcac catcgtcaaa ccccaggagt cagcccaccg gaggatggag ccccctgcct
     1441 ccaaggtccc cgaggtgccc actgcccctg ccaccgacgc agcccccaag agggtggaga
     1501 tccagatgcc caagcctgct gaggcgccca ccgcccccag cccagcccag accttggaaa
     1561 attcagagcc tgcccctgtg tctcagctgc agagcaggct ggagcccaag ccccagcccc
     1621 ctgtggctga ggctacaccc cggagccagg aggccactga ggcggctccc agctgcgttg
     1681 gcgacatggc cgacaccccc agagatgccg ggctcaagca ggcgcctgca tcacggaacg
     1741 agaaggcccc ggtggacttc ggctacgtgg ggattgactc catcctggag cagatgcgcc
     1801 ggaaggccat gaagcagggc ttcgagttca acatcatggt ggtcgggcag agcggcttgg
     1861 gtaaatccac cttaatcaac accctcttca aatccaaaat cagccggaag tcggtgcagc
     1921 ccacctcaga ggagcgcatc cccaagacca tcgagatcaa gtccatcacg cacgatattg
     1981 aggagaaagg cgtccggatg aagctgacag tgattgacac accagggttc ggggaccaca
     2041 tcaacaacga gaactgctgg cagcccatca tgaagttcat caatgaccag tacgagaaat
     2101 acctgcagga ggaggtcaac atcaaccgca agaagcgcat cccggacacc cgcgtccact
     2161 gctgcctcta cttcatcccc gccaccggcc actccctcag gcccctggac atcgagttta
     2221 tgaaacgcct gagcaaggtg gtcaacatcg tccctgtcat cgccaaggcg gacacactca
     2281 ccctggagga gagggtccac ttcaaacagc ggatcaccgc agacctgctg tccaacggca
     2341 tcgacgtgta cccccagaag gaatttgatg aggactcgga ggaccggctg gtgaacgaga
     2401 agttccggga gatgatccca tttgctgtgg tgggcagtga ccacgagtac caggtcaacg
     2461 gcaagaggat ccttgggagg aagaccaagt ggggtaccat cgaagttgaa aacaccacac
     2521 actgtgagtt tgcctacctg cgggaccttc tcatcaggac gcacatgcag aacatcaagg
     2581 acatcaccag cagcatccac ttcgaggcgt accgtgtgaa gcgcctcaac gagggcagca
     2641 gcgccatggc caacggcgtg gaggagaagg agccagaagc cccagagatg atcgatggtg
     2701 gcggtggctc tggaggtggt gggtcctccg gagagaagcg cgaccacatg gtgctgctgg
     2761 agtacgtgac cgccgccggc atcaccgacg cctcctaatc tagagggccc gtttaaaccc
     2821 gctgatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc ccctcccccg
     2881 tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa aatgaggaaa
     2941 ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg gggtggggtg gggcaggaca
     3001 gcaaggggga ggattgggaa gacaatagca ggcatgctgg ggatgcggtg ggctctatgg
     3061 cttctgaggc ggaaagaacc agctggggct ctagggggta tccccacgcg ccctgtagcg
     3121 gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt gaccgctaca cttgccagcg
     3181 ccctagcgcc cgctcctttc gctttcttcc cttcctttct cgccacgttc gccggctttc
     3241 cccgtcaagc tctaaatcgg ggcatccctt tagggttccg atttagtgct ttacggcacc
     3301 tcgaccccaa aaaacttgat tagggtgatg gttcacgtag tgggccatcg ccctgataga
     3361 cggtttttcg ccctttgacg ttggagtcca cgttctttaa tagtggactc ttgttccaaa
     3421 ctggaacaac actcaaccct atctcggtct attcttttga tttataaggg attttgggga
     3481 tttcggccta ttggttaaaa aatgagctga tttaacaaaa atttaacgcg aattaattct
     3541 gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc tccccaggca ggcagaagta
     3601 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     3661 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     3721 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     3781 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     3841 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     3901 ccattttcgg atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat
     3961 agtataatac gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt
     4021 gctcaccgcg cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc
     4081 ccgggacttc gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat
     4141 cagcgcggtc caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg
     4201 cctggacgag ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc
     4261 cgggccggcc atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga
     4321 cccggccggc aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga
     4381 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4441 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4501 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4561 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4621 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc
     4681 tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg
     4741 taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc
     4801 cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg
     4861 gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc
     4921 ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac
     4981 agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa
     5041 ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca
     5101 caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc
     5161 gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata
     5221 cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac gctgtaggta
     5281 tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca
     5341 gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga
     5401 cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg
     5461 tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga cagtatttgg
     5521 tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg
     5581 caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag
     5641 aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa
     5701 cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat
     5761 ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc
     5821 tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     5881 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     5941 tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc
     6001 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     6061 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     6121 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     6181 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     6241 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     6301 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     6361 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     6421 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     6481 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     6541 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     6601 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     6661 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     6721 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     6781 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtc
//
