LOCUS       62056_21630        6214 bp DNA     circular SYN 25-JAN-2021
DEFINITION  MCherry and eGFP expression vector pTTC-Hsap-WT, complete sequence.
ACCESSION   MN232103
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6214)
  AUTHORS   Boldyreva LV, Yarinich LA, Kozhevnikova EN, Ivankin AV, Lebedev MO, 
            Pindyurin AV.
  TITLE     Fine gene expression regulation by minor sequence variations 
            downstream of the polyadenylation signal
  JOURNAL   Mol Biol Rep (2021) In press
REFERENCE   2  (bases 1 to 6214)
  AUTHORS   Boldyreva LV, Ivankin AV, Kozhevnikova EN, Pindyurin AV.
  TITLE     Direct Submission
  JOURNAL   Submitted (25-JUL-2019) Department of the Regulation of Genetic 
            Processes, Institute of Molecular and Cellular Biology, Lavrentyev 
            ave. 8/2, Novosibirsk, Novosibirsk Region 630090, Russia
REFERENCE   3  (bases 1 to 6214)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol Biol 
            Rep (2021) In press"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Submitted 
            (25-JUL-2019) Department of the Regulation of Genetic Processes, 
            Institute of Molecular and Cellular Biology, Lavrentyev ave. 8/2, 
            Novosibirsk, Novosibirsk Region 630090, Russia"
FEATURES             Location/Qualifiers
     source          1..6214
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    838..859
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        874..904
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    912..928
                     /label=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)."
     primer_bind     936..952
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(989..1122)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1132..1836)
                     /label=mCherry
                     /note="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
     promoter        complement(1913..1931)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1976..2179)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     enhancer        complement(2180..2483)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2558..3068
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             3078..3794
                     /label=EGFP
                     /note="enhanced GFP"
     regulatory      3890..3906
                     /label=human neuropilin 1 (NRP1) polyA signal
                     /note="human neuropilin 1 (NRP1) polyA signal"
                     /regulatory_class="polyA_signal_sequence"
     misc_feature    3913..3979
                     /note="piggyBac transposon 3' terminal repeat sequence
                     together with the adjacent TTAA target site"
     promoter        complement(4026..4044)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(4065..4081)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4089..4105)
                     /label=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(4113..4143)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4158..4179)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4467..5055)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5229..6086)
                     /label=AmpR
                     /note="beta-lactamase"
     promoter        complement(6087..6191)
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tggggcgcgc
      661 cattctagat gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
      721 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
      781 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
      841 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
      901 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
      961 cgccaagctc tagctagagg tcgacggtat acagacatga taagatacat tgatgagttt
     1021 ggacaaacca caactagaat gcagtgaaaa aaatgcttta tttgtgaaat ttgtgatgct
     1081 attgctttat ttgtaaccat tataagctgc aataaacaag ttgatatctt acttgtacag
     1141 ctcgtccatg ccgccggtgg agtggcggcc ctcggcgcgt tcgtactgtt ccacgatggt
     1201 gtagtcctcg ttgtgggagg tgatgtccaa cttgatgttg acgttgtagg cgccgggcag
     1261 ctgcacgggc ttcttggcct tgtaggtggt cttgacctca gcgtcgtagt ggccgccgtc
     1321 cttcagcttc agcctctgct tgatctcgcc cttcagggcg ccgtcctcgg ggtacatccg
     1381 ctcggaggag gcctcccagc ccatggtctt cttctgcatt acggggccgt cggaggggaa
     1441 gttggtgccg cgcagcttca ccttgtagat gaactcgccg tcctgcaggg aggagtcctg
     1501 ggtcacggtc accacgccgc cgtcctcgaa gttcatcacg cgctcccact tgaagccctc
     1561 ggggaaggac agcttcaagt agtcggggat gtcggcgggg tgcttcacgt aggccttgga
     1621 gccgtacatg aactgagggg acaggatgtc ccaggcgaag ggcagggggc cacccttggt
     1681 caccttcagc ttggcggtct gggtgccctc gtaggggcgg ccctcgccct cgccctcgat
     1741 ctcgaactcg tggccgttca cggagccctc catgtgcacc ttgaagcgca tgaactcctt
     1801 gatgatggcc atgttatcct cctcgccctt gctcacccca agctttcttc gtcgctgtct
     1861 ccgcttcttc ctgccataca tggatccgag ctcggtacca agcgagggtc tccctatagt
     1921 gagtcgtatt aatttcgata agccagtaag cagtgggttc tctagttagc cagagagctc
     1981 tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt
     2041 tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc ccattgacgt
     2101 caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc ccattgatgt
     2161 actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg tactgccaag
     2221 taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat ttaccgtcat
     2281 tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc aagtgggcag
     2341 tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc gttactatgg
     2401 gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc
     2461 catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact aatgaccccg
     2521 taattgatta ctattaatgc tagcagatct accggtcggg gttggggttg cgccttttcc
     2581 aaggcagccc tgggtttgcg cagggacgcg gctgctctgg gcgtggttcc gggaaacgca
     2641 gcggcgccga ccctgggtct cgcacattct tcacgtccgt tcgcagcgtc acccggatct
     2701 tcgccgctac ccttgtgggc cccccggcga cgcttcctgc tccgccccta agtcgggaag
     2761 gttccttgcg gttcgcggcg tgccggacgt gacaaacgga agccgcacgt ctcactagta
     2821 ccctcgcaga cggacagcgc cagggagcaa tggcagcgcg tcgaccgcga tgggctgtgg
     2881 ccaatagcgg ctgctcagca gggcgcgccg agagcagcgg ccgggaaggg gcggtgcggg
     2941 aggcggggtg tggggcggta gtgtgggccc tgttcctgcc cgcgcggtgt tccgcattct
     3001 gcaagcctcc ggagcgcacg tcggcagtcg gctccctcgt tgaccgaatc accgacctct
     3061 ctccccagcc cgggaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3121 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3181 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3241 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3301 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3361 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     3421 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     3481 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     3541 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     3601 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     3661 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     3721 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     3781 acgagctgta caagtaagaa ttcgcggccg catacgattt aggtgacact cgaggatcga
     3841 gatttaaatg agttgtggcc ggcccttgtg actgggaaaa ccctggcgta aataaaatac
     3901 gaaatgacta gtcatgcgtc aattttacgc atgattatct ttaacgtacg tcacaatatg
     3961 attatctttc tagggttaat ctagtatacg cgtatgcggc cgcttaatta atccagcttt
     4021 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     4081 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     4141 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     4201 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     4261 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     4321 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     4381 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     4441 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     4501 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     4561 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     4621 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     4681 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     4741 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     4801 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     4861 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     4921 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     4981 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     5041 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     5101 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     5161 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     5221 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     5281 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     5341 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     5401 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     5461 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     5521 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     5581 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     5641 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     5701 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     5761 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     5821 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     5881 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     5941 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6001 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     6061 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     6121 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     6181 ggggttccgc gcacatttcc ccgaaaagtg ccac
//
