LOCUS       Exported               10951 bp ds-DNA    circular SYN 15-11ÔÂ-2018
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    pCAG-C4-GFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10951)
  AUTHORS   gen
  TITLE     Direct Submission
  JOURNAL   Exported 2018Äê11ÔÂ15ÈÕ from SnapGene 2.3.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10951
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        4..383
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /note="chicken beta-actin promoter"
     CDS             7168..7887
                     /codon_start=1
                     /product="enhanced GFP"
                     /note="EGFP"
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_site      8039..8094
                     /note="beta-globin poly(A) signal"
                     /note="rabbit beta-globin polyadenylation signal"
     primer_bind     complement(8454..8470)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    8478..8494
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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(8502..8532)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    8547..8568
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        8627..8822
                     /note="SV40 promoter"
                     /note="SV40 early promoter"
     rep_origin      8673..8808
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     polyA_signal    8828..8962
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(9201..9789)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(9960..10820)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(10821..10925)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gcccccggag cgccggcggc
     1321 tgtcgaggcg cggcgagccg cagccattgc cttttatggt aatcgtgcga gagggcgcag
     1381 ggacttcctt tgtcccaaat ctgtgcggag ccgaaatctg ggaggcgccg ccgcaccccc
     1441 tctagcgggc gcggggcgaa gcggtgcggc gccggcagga aggaaatggg cggggagggc
     1501 cttcgtgcgt cgccgcgccg ccgtcccctt ctccctctcc agcctcgggg ctgtccgcgg
     1561 ggggacggct gccttcgggg gggacggggc agggcggggt tcggcttctg gcgtgtgacc
     1621 ggcggctcta gagcctctgc taaccatgtt catgccttct tctttttcct acagctcctg
     1681 ggcaacgtgc tggttattgt gctgtctcat cattttggca aagaattctg cagtcgacgg
     1741 taccgcgggc ccgggatcca ccggtagaag ggagcagacg tctgatttca gtcagaccag
     1801 acaggtctga cctttcctgg atcctccagc catgcggctc ctctgggggc tggcctgggt
     1861 gttcagcttc tgtgcctcat ccctgcagaa gcccaggttg ctcctgtttt ccccttctgt
     1921 ggttaatttg gggacccccc tgtcggtggg ggtacagctc ctggatgccc ctccaggaca
     1981 ggaggtaaaa ggatcagtgt tcctcagaaa cccaaagggt ggttcctgct ccccaaagaa
     2041 ggactttaag ctgagctcgg gagatgactt tgtgctgctc agccttgagg tcccactgga
     2101 agatgtgagg agctgtggcc tctttgacct gcgcagagcc ccccacatcc agctggtagc
     2161 tcagtctccg tggctaagga acacagcttt caaagccaca gagactcagg gtgtcaactt
     2221 gctcttctct tcccgacgag gccacatctt tgtgcagacc gatcagccta tctataatcc
     2281 ggggcagcgg gttcgttatc gggtctttgc actggatcaa aagatgcgcc catccactga
     2341 tttcctcacc atcacagtgg agaactccca tggcctccgt gtactcaaga aggagatatt
     2401 tacttccaca tccatcttcc aagatgcctt aactcccatg gacatctcag agcctgggac
     2461 ctggaagatc tcagctaggt tctcagatgg actggagtcc aataggagca cccactttga
     2521 agtgaagaag tatgtccttc ccaacttcga ggtgaagatt actccttgga agccatatat
     2581 cctgatggtg cccagcaaca gtgatgaaat ccaattagac atccaggcca ggtacatcta
     2641 tgggaagccc gtgcagggcg tggcatacac acggtttgcg ctcatggatg agcaagggaa
     2701 gaggactttc cttcggggcc tagagacgca ggccaagttg gtggaaggcc ggacccacat
     2761 ttccatctca aaggaccagt tccaggctgc cctggataaa atcaatattg gggtcagaga
     2821 cctggagggg ctgcgtctct atgctgctac agctgtcatc gagtctccag gaggagagat
     2881 ggaggaggca gaactcacgt cctggcgctt tgtatcatct gccttctcct tggatctcag
     2941 ccgcactaag aggcatcttg tgcctggggc ccacttcctg ctgcaggctc tggtccaaga
     3001 aatgtcaggc tctgaagcct ccaacgttcc tgttaaagtc tctgccacat tggtgtcagg
     3061 ctctgactcc caagtccttg acattcaaca gagcaccaat ggaattggcc aagtcagcat
     3121 ttccttcccc atcccaccaa ctgtcacaga acttcgactc ttggtgtctg cgggctccct
     3181 ctacccagcc atagccaggc tcaccgtgca agccccacct tcaagaggca ctggctttct
     3241 ttctattgag ccactagacc ctcggtcccc tagtgtgggg gacaccttta tcctaaacct
     3301 tcaacctgtg ggcatccctg cacctacctt ctctcattac tactacatga tcatctccag
     3361 aggccagatc atggctatgg gtcgggagcc ccggaagact gtgacctccg tctctgtgtt
     3421 ggtggaccat cagctggctc cctcgttcta ctttgtggct tacttctatc accaaggaca
     3481 cccggtggcc aactctctgc tcatcaacat ccaatccagg gactgtgagg gcaagctgca
     3541 attgaaggtg gatggtgcca aggagtatcg taatgcggac atgatgaagc tccgaattca
     3601 aactgactcc aaagccctgg tggcactggg agctgtggac acggctctgt atgctgtggg
     3661 tggtcggtct cacaaacccc tcgacatgag caaggtcttt gaagtaatca acagctacaa
     3721 tgttggctgt ggtcctggag gtggggatga tgcccttcag gtgttccagg atgctggtct
     3781 ggccttttct gatggtgatc gactaactca aaccagagag gacctgagct gtcccaagga
     3841 gaagaaaagt cggcaaaaga gaaatgttaa cttccagaag gctgtcagtg agaagttggg
     3901 ccagtattct tctccagatg ccaagcgctg ctgccaagac gggatgacga agctgcccat
     3961 gaagcgtacc tgtgagcagc gggctgcccg tgtgcctcag caggcctgcc gtgagccctt
     4021 cttgtcctgt tgcaagtttg ctgaggacct tcgcaggaac cagaccagga gccaggcaca
     4081 ccttgcccga aacaaccaca acatgctgca ggaggaagac ttgatagatg aagacgacat
     4141 tcttgtgcgc acctccttcc cagagaactg gctctggaga gtggaacctg tagacagctc
     4201 caaactgttg acagtgtggc ttcctgattc tatgaccaca tgggagattc atggtgtgag
     4261 cctgtccaaa agcaaaggtc tgtgtgtagc caagccaact cgtgttcgag tgttcagaaa
     4321 attccacctg cacctgcgcc tgcccatctc catccgccgc tttgagcagt ttgaattacg
     4381 gcctgttctt tacaactatc tgaatgatga tgtggctgtg agtgtccatg tgaccccagt
     4441 tgaggggctg tgcctggctg gtggtggaat gatggcccag caggtgacag tgccagcagg
     4501 ttctgcccgg cctgtggcct tctctgtggt acccacagct gctgccaacg tgcccctgaa
     4561 ggtggtggct cgaggggttt ttgatttagg ggatgctgtg tctaagattc tccaaattga
     4621 gaaggaagga gccatccaca gagaagagtt agtctacaac ctcgaccccc taaataacct
     4681 gggtcggact ttggagattc ctggcagctc ggatcccaac atcgtccctg acggagactt
     4741 cagcagctta gtcagggtta cagcctcgga acccttggag actatgggtt ctgaaggtgc
     4801 cttgtcccca ggaggtgtgg cctccctcct gaggcttccc cagggctgtg cagagcaaac
     4861 catgatctat ttggctccta ccctgactgc ttccaactac ctggacagga cagaacagtg
     4921 gagcaaactg tcccctgaaa caaaggacca tgctgtggat ctgatccaaa aaggatacat
     4981 gaggatccag cagtttcgga agaatgatgg ctcctttggg gcttggttac accgggacag
     5041 cagcacctgg ctgactgcct tcgtgctgaa gattctgagt ttggcccagg aacaggtggg
     5101 caactccccg gagaagctgc aggagacggc tagctggctg ctggcccagc agctgggtga
     5161 tggctccttc cacgacccat gtccagtcat ccacagagca atgcaggggg gcttggtggg
     5221 gtctgacgag acagtggcac tgaccgcctt tgtggtcatt gcccttcacc atgggttgga
     5281 cgtcttccag gatgacgatg cgaagcagct gaagaacaga gtggaagcct ccatcaccaa
     5341 ggcaaactca ttcttggggc agaaggcaag tgctgggctc ctgggtgccc acgccgccgc
     5401 tatcacagcc tatgccctta cgctgaccaa ggcctcggag gacctgcaga atgttgccca
     5461 caacagcctg atggccatgg ctgaggaaac aggggaacac ctctactggg gcttagtcct
     5521 tggctctcag gacaaagttg tgttgcgccc cacagccccc cgtagcccta cagaacctgt
     5581 gccccaggcc ccagccttgt ggatcgaaac cacagcctat gccctgctcc acctgcttct
     5641 gcgtgaggga aagggaaaaa tggctgacaa ggctgcatcc tggctcaccc accagggaag
     5701 cttccatggg gcattccgca gtacccagga cactgtggtc accctggatg ccctgtctgc
     5761 ctactggatc gcttcgcaca ccactgagga gaaagcactg aacgtgacgc tcagctccat
     5821 gggccgcaat gggcttaaga cccacgggct acacttgaac aaccaccaag tcaagggcct
     5881 ggaggaggag ctaaagttct ccctgggcag cacaatcagt gtcaaggtgg aaggaaacag
     5941 caaaggcacc ttgaagatcc ttcgtaccta caatgtcctg gacatgaaga acaccacatg
     6001 ccaggacctt cagatagaag tgaaggtcac aggcgctgtg gaatatgcat gggatgccaa
     6061 tgaagactac gaagactact atgacatgcc agccgcagat gaccccagcg tccccttgca
     6121 gcctgtcacg cccctgcagc tttttgaggg tcgtaggagc cgccgcagga gggaggcccc
     6181 caaggtggct gaagagcggg agtccagagt tcagtacact gtgtgtatct ggcgaaatgg
     6241 caagctgggg ctgtctggca tggccatcgc agacatcacc ctcctaagtg gattccacgc
     6301 cctgagggct gacctggaga agctgacctc cctctctgac cgttacgtga gtcactttga
     6361 gactgacggg ccccatgtcc tgttgtactt tgactcggtc cctaccaccc gggagtgtgt
     6421 gggcttcgga gcctcgcagg aggtggttgt gggactggtg cagccatcca gtgctgtcct
     6481 gtatgactac tacagccctg atcacaagtg ctctgtgttt tatgctgcac ccaccaagag
     6541 ccagctcctg gccacactgt gctctggaga tgtatgccag tgcgctgagg ggaagtgccc
     6601 tcgactgcta aggtcactgg agcgaagggt ggaggacaag gatggctacc ggatgaggtt
     6661 cgcctgctat tatccccgag tggagtatgg cttcacggtt aaggttcttc gagaagatgg
     6721 cagagctgcc ttccgtctct ttgagtccaa gatcacccaa gtcctgcatt tcagaaagga
     6781 caccatggcc tccataggtc agacccgcaa cttcctgagc cgggcctctt gccgccttcg
     6841 tttggagcct aacaaagagt acttgatcat ggggatggac ggggaaacca gtgacaacaa
     6901 gggagacccc cagtacttgc tggactcaaa tacctggatt gaggagatgc cttcagaaca
     6961 aatgtgcaag agcacccgcc atcgggcagc ctgtttccag ctcaaagact tcctgatgga
     7021 gttcagcagc cgggggtgcc aggtgtgagg ccttaggact ctggctctct gagctcagct
     7081 cagggttagg gcctcactgg attagaggct ctgctctaca gggtaaataa aagaaaagct
     7141 ttttgacagc caaaaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg
     7201 gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc
     7261 ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc
     7321 ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc
     7381 ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa
     7441 ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc
     7501 gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc
     7561 aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc
     7621 tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac
     7681 atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac
     7741 ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac
     7801 cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact
     7861 ctcggcatgg acgagctgta caagtaaagc ggccgcactc ctcaggtgca ggctgcctat
     7921 cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt
     7981 ccctctgcca aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa
     8041 taaaggaaat ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga
     8101 aggacatatg ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg
     8161 gcaacatatg cccatatgct ggctgccatg aacaaaggtg gctataaaga ggtcatcagt
     8221 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     8281 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     8341 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     8401 ctcttctctt atgaagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     8461 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     8521 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     8581 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     8641 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     8701 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     8761 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     8821 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     8881 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     8941 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     9001 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     9061 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     9121 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     9181 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     9241 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     9301 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     9361 gcctttctcc cttcgggaag cgtggcgctt tctcaatgct cacgctgtag gtatctcagt
     9421 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     9481 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     9541 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     9601 gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc
     9661 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     9721 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     9781 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     9841 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     9901 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     9961 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
    10021 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
    10081 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
    10141 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
    10201 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
    10261 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
    10321 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
    10381 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
    10441 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
    10501 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
    10561 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
    10621 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
    10681 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
    10741 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
    10801 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
    10861 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
    10921 ccgcgcacat ttccccgaaa agtgccacct g
//
