LOCUS       Exported                8851 bp ds-DNA     circular SYN 04-JUN-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pFRTpsiCHECK_SEAP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8851)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Jun 8, 2020 from SnapGene Viewer 2.7.3
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8851
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        73..430
                     /note="SV40 promoter"
                     /note="SV40 enhancer and early promoter"
     rep_origin      281..416
                     /note="SV40 ori"
                     /note="SV40 origin of replication"
     intron          500..632
                     /note="chimeric intron"
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        677..695
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             701..2203
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /note="SEAP"
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="MLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQP
                     AQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDK
                     HVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVV
                     TTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRK
                     YMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTH
                     LMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESR
                     AYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKA
                     RDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFA
                     RGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     polyA_signal    2261..2309
                     /note="synthetic polyadenylation signal"
     promoter        2318..3069
                     /note="HSV TK promoter"
                     /note="herpes simplex virus thymidine kinase promoter"
     misc_feature    3105..4757
                     /note="Synthetic Firefly luciferase"
     polyA_signal    4809..5033
                     /note="bGH poly(A) signal"
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    5317..5364
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FRT"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     CDS             5372..6394
                     /codon_start=1
                     /gene="aph(4)-Ia"
                     /product="aminoglycoside phosphotransferase from E. coli"
                     /note="HygR"
                     /note="confers resistance to hygromycin"
                     /translation="KKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRGY
                     VLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLPE
                     TELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQT
                     VMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMFG
                     DSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDGN
                     FDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAKE
                     "
     polyA_signal    6524..6645
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6694..6710)
                     /note="M13 rev"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    6718..6734
                     /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(6742..6772)
                     /note="lac promoter"
                     /note="promoter for the E. coli lac operon"
     protein_bind    6787..6808
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding site"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(7096..7684)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7855..8715)
                     /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(8716..8820)
                     /gene="bla"
                     /note="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctgcg cagcaccatg gcctgaaata acctctgaaa gaggaacttg
       61 gttaggtacc ttctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
      121 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
      181 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
      241 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
      301 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
      361 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
      421 cttttgcaaa aagcttgatt cttctgacac aacagtctcg aacttaagct gcagaagttg
      481 gtcgtgaggc actgggcagg taagtatcaa ggttacaaga caggtttaag gagaccaata
      541 gaaactgggc ttgtcgagac agagaagact cttgcgtttc tgataggcac ctattggtct
      601 tactgacatc cactttgcct ttctctccac aggtgtccac tcccagttca attacagctc
      661 ttaaggctag agtacttaat acgactcact ataggctagc atgctgctgc tgctgctgct
      721 gctgggcctg aggctacagc tctccctggg catcatccca gttgaggagg agaacccgga
      781 cttctggaac cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca
      841 gacagccgcc aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac
      901 agctgccagg atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc
      961 catggaccgc ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc
     1021 agacagtgga gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat
     1081 tggcttgagt gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat
     1141 ctccgtgatg aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg
     1201 agtgcagcac gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc
     1261 ggacgccgac gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct
     1321 catctccaac atggacattg acgtgatcct aggtggaggc cgaaagtaca tgtttcgcat
     1381 gggaacccca gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg
     1441 gaagaatctg gtgcaggaat ggctggcgaa gcgccagggt gcccggtatg tgtggaaccg
     1501 cactgagctc atgcaggctt ccctggaccc gtctgtgacc catctcatgg gtctctttga
     1561 gcctggagac atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga
     1621 gatgacagag gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt
     1681 ggagggtggt cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga
     1741 gacgatcatg ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac
     1801 gctgagcctc gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg
     1861 agggagctcc atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt
     1921 cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac
     1981 cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga
     2041 gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca
     2101 cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc
     2161 ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggtta
     2221 ctctagagtc ggggcggccg gctaggcggc cgctggccgc aataaaatat ctttattttc
     2281 attacatctg tgtgttggtt ttttgtgtga ggatctaaat gagtcttcgg acctcgcggg
     2341 ggccgcttaa gcggtggtta gggtttgtct gacgcggggg gagggggaag gaacgaaaca
     2401 ctctcattcg gaggcggctc ggggtttggt cttggtggcc acgggcacgc agaagagcgc
     2461 cgcgatcctc ttaagcaccc ccccgccctc cgtggaggcg ggggtttggt cggcgggtgg
     2521 taactggcgg gccgctgact cgggcgggtc gcgcgcccca gagtgtgacc ttttcggtct
     2581 gctcgcagac ccccgggcgg cgccgccgcg gcggcgacgg gctcgctggg tcctaggctc
     2641 catggggacc gtatacgtgg acaggctctg gagcatccgc acgactgcgg tgatattacc
     2701 ggagaccttc tgcgggacga gccgggtcac gcggctgacg cggagcgtcc gttgggcgac
     2761 aaacaccagg acggggcaca ggtacactat cttgtcaccc ggaggcgcga gggactgcag
     2821 gagcttcagg gagtggcgca gctgcttcat ccccgtggcc cgttgctcgc gtttgctggc
     2881 ggtgtccccg gaagaaatat atttgcatgt ctttagttct atgatgacac aaaccccgcc
     2941 cagcgtcttg tcattggcga attcgaacac gcagatgcag tcggggcggc gcggtcccag
     3001 gtccacttcg catattaagg tgacgcgtgt ggcctcgaac accgagcgac cctgcagcga
     3061 cccgcttaaa agcttggcat tccggtactg ttggtaaagc caccatggcc gatgctaaga
     3121 acattaagaa gggccctgct cccttctacc ctctggagga tggcaccgct ggcgagcagc
     3181 tgcacaaggc catgaagagg tatgccctgg tgcctggcac cattgccttc accgatgccc
     3241 acattgaggt ggacatcacc tatgccgagt acttcgagat gtctgtgcgc ctggccgagg
     3301 ccatgaagag gtacggcctg aacaccaacc accgcatcgt ggtgtgctct gagaactctc
     3361 tgcagttctt catgccagtg ctgggcgccc tgttcatcgg agtggccgtg gcccctgcta
     3421 acgacattta caacgagcgc gagctgctga acagcatggg catttctcag cctaccgtgg
     3481 tgttcgtgtc taagaagggc ctgcagaaga tcctgaacgt gcagaagaag ctgcctatca
     3541 tccagaagat catcatcatg gactctaaga ccgactacca gggcttccag agcatgtaca
     3601 cattcgtgac atctcatctg cctcctggct tcaacgagta cgacttcgtg ccagagtctt
     3661 tcgacaggga caaaaccatt gccctgatca tgaacagctc tgggtctacc ggcctgccta
     3721 agggcgtggc cctgcctcat cgcaccgcct gtgtgcgctt ctctcacgcc cgcgacccta
     3781 ttttcggcaa ccagatcatc cccgacaccg ctattctgag cgtggtgcca ttccaccacg
     3841 gcttcggcat gttcaccacc ctgggctacc tgatttgcgg ctttcgggtg gtgctgatgt
     3901 accgcttcga ggaggagctg ttcctgcgca gcctgcaaga ctacaaaatt cagtctgccc
     3961 tgctggtgcc aaccctgttc agcttcttcg ctaagagcac cctgatcgac aagtacgacc
     4021 tgtctaacct gcacgagatt gcctctggcg gcgccccact gtctaaggag gtgggcgaag
     4081 ccgtggccaa gcgctttcat ctgccaggca tccgccaggg ctacggcctg accgagacaa
     4141 ccagcgccat tctgattacc ccagagggcg acgacaagcc tggcgccgtg ggcaaggtgg
     4201 tgccattctt cgaggccaag gtggtggacc tggacaccgg caagaccctg ggagtgaacc
     4261 agcgcggcga gctgtgtgtg cgcggcccta tgattatgtc cggctacgtg aataaccctg
     4321 aggccacaaa cgccctgatc gacaaggacg gctggctgca ctctggcgac attgcctact
     4381 gggacgagga cgagcacttc ttcatcgtgg accgcctgaa gtctctgatc aagtacaagg
     4441 gctaccaggt ggccccagcc gagctggagt ctatcctgct gcagcaccct aacattttcg
     4501 acgccggagt ggccggcctg cccgacgacg atgccggcga gctgcctgcc gccgtcgtcg
     4561 tgctggaaca cggcaagacc atgaccgaga aggagatcgt ggactatgtg gccagccagg
     4621 tgacaaccgc caagaagctg cgcggcggag tggtgttcgt ggacgaggtg cccaagggcc
     4681 tgaccggcaa gctggacgcc cgcaagatcc gcgagatcct gatcaaggct aagaaaggcg
     4741 gcaagatcgc cgtgtaataa ttctagtcga gtctagaggg cccgtttaaa cccgctgatc
     4801 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc
     4861 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc
     4921 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg
     4981 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga
     5041 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt
     5101 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc
     5161 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca
     5221 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc
     5281 caaaaaactt gattagggtg atggttcacg tacctagaag ttcctattcc gaagttccta
     5341 ttctctagaa agtataggaa cttccttggc caaaaagcct gaactcaccg cgacgtctgt
     5401 cgagaagttt ctgatcgaaa agttcgacag cgtctccgac ctgatgcagc tctcggaggg
     5461 cgaagaatct cgtgctttca gcttcgatgt aggagggcgt ggatatgtcc tgcgggtaaa
     5521 tagctgcgcc gatggtttct acaaagatcg ttatgtttat cggcactttg catcggccgc
     5581 gctcccgatt ccggaagtgc ttgacattgg ggaattcagc gagagcctga cctattgcat
     5641 ctcccgccgt gcacagggtg tcacgttgca agacctgcct gaaaccgaac tgcccgctgt
     5701 tctgcagccg gtcgcggagg ccatggatgc gatcgctgcg gccgatctta gccagacgag
     5761 cgggttcggc ccattcggac cgcaaggaat cggtcaatac actacatggc gtgatttcat
     5821 atgcgcgatt gctgatcccc atgtgtatca ctggcaaact gtgatggacg acaccgtcag
     5881 tgcgtccgtc gcgcaggctc tcgatgagct gatgctttgg gccgaggact gccccgaagt
     5941 ccggcacctc gtgcacgcgg atttcggctc caacaatgtc ctgacggaca atggccgcat
     6001 aacagcggtc attgactgga gcgaggcgat gttcggggat tcccaatacg aggtcgccaa
     6061 catcttcttc tggaggccgt ggttggcttg tatggagcag cagacgcgct acttcgagcg
     6121 gaggcatccg gagcttgcag gatcgccgcg gctccgggcg tatatgctcc gcattggtct
     6181 tgaccaactc tatcagagct tggttgacgg caatttcgat gatgcagctt gggcgcaggg
     6241 tcgatgcgac gcaatcgtcc gatccggagc cgggactgtc gggcgtacac aaatcgcccg
     6301 cagaagcgcg gccgtctgga ccgatggctg tgtagaagta ctcgccgata gtggaaaccg
     6361 acgccccagc actcgtccga gggcaaagga atagcacgta ctacgagatt tcgattccac
     6421 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     6481 cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt ttattgcagc
     6541 ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc
     6601 actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg tctgtatacc
     6661 gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     6721 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     6781 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     6841 gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga gaggcggttt
     6901 gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
     6961 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga
     7021 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
     7081 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
     7141 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
     7201 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
     7261 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
     7321 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg
     7381 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
     7441 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
     7501 cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta tctgcgctct
     7561 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
     7621 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
     7681 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg
     7741 ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta
     7801 aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca
     7861 atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc
     7921 ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc
     7981 tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc
     8041 agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat
     8101 taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt
     8161 tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc
     8221 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
     8281 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
     8341 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
     8401 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
     8461 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
     8521 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
     8581 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
     8641 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
     8701 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
     8761 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
     8821 cacatttccc cgaaaagtgc cacctgacgt c
//
