LOCUS       Exported                7185 bp ds-DNA     circular SYN 16-MAR-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7185)
  AUTHORS   aneuss
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Mar 16, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7185
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     misc_feature    1..25
                     /label=MCS
                     /note="multiple cloning site"
                     /note="color: #99ccff"
     promoter        30..607
                     /label=CMV immearly promoter
                     /note="color: #ffffff"
     misc_feature    66..369
                     /label=CMV_enhancer
                     /note="color: #008080"
     misc_feature    370..573
                     /label=CMV_promoter
                     /note="color: #008080; direction: RIGHT"
     misc_feature    613..616
                     /label=Ligation
                     /label=nonstandard type:  ligation
                     /note="color: #a6acb3"
     misc_feature    625..634
                     /label=Kozak-Sequenz
                     /note="color: #ff99cc"
     CDS             631..1350
                     /codon_start=1
                     /label=ORF frame 1
                     /note="color: #993366"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             631..1350
                     /codon_start=1
                     /label=EGFP
                     /note="color: #00ff00"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1374..1485
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
                     /note="color: #a6acb3"
     promoter        1608..1937
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
                     /note="color: #ffffff; direction: RIGHT"
     rep_origin      1788..1923
                     /label=SV40 ori
                     /note="SV40 origin of replication"
                     /note="color: #ffff00"
     promoter        1985..2032
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
                     /note="color: #ffffff; direction: RIGHT"
     CDS             2051..2650
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /note="color: #ccffcc"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    2780..2901
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     promoter        2966..3482
                     /label=CMV_immearly Promotor
                     /note="color: #ffffff"
     promoter        3440..3509
                     /label=CMV_Promoter
                     /note="color: #ffffff"
     CDS             3537..4214
                     /codon_start=1
                     /label=mPlum
                     /note="color: #ff0000"
                     /translation="MVSKGEEVIKEFMRFKEHMEGSVNGHEFEIEGEGEGRPYEGTQTA
                     RLKVTKGGPLPFAWDILSPQIMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGG
                     VVTVTQDSSLQDGEFIYKVKVRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALKGEMK
                     MRLRLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRH
                     STGA"
     terminator      4227..4455
                     /label=SV40_Terminator
                     /note="color: #ffffff"
     CDS             complement(4509..5369)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1: 4509 .. 5300 / #ccffcc
                        2: 5301 .. 5369 / #ccffcc / signal sequence
                     Cleavage site after base 5300"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5633..6221
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     protein_bind    6963..6979
                     /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)."
                     /note="color: #31849b"
     misc_feature    7094..7185
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
                     /note="color: #a6acb3"
ORIGIN
        1 actagtgaat tcggtaccga gctcctcgag gggtcattag ttcatagccc atatatggag
       61 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      121 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      181 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      241 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      301 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      361 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      421 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      481 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      541 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      601 tactggctta tggatccacc ggtcgccacc atggtgagca agggcgagga gctgttcacc
      661 ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg
      721 tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc
      781 accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag
      841 tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc
      901 gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc
      961 gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac
     1021 ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac
     1081 gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac
     1141 aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc
     1201 gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa
     1261 gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc
     1321 actctcggca tggacgagct gtacaagtaa agcggccgca aaatcagcct cgactgtgcc
     1381 ttctagttgc cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg
     1441 tgccactccc actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct
     1501 atctcggtct attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa
     1561 aatgagctga tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag
     1621 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     1681 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     1741 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1801 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1861 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1921 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc
     1981 acgtgttgac aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag
     2041 gaactaaacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     2101 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     2161 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     2221 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     2281 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     2341 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     2401 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     2461 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     2521 cgccttcctg gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     2581 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     2641 cggtgcctga cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg
     2701 cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct
     2761 ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa
     2821 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     2881 caaactcatc aatgtatctt atcatgtctg tatagctcgt agatttaatt aaacgcgtgc
     2941 gatcgcaagg ccgtcaaggc cgcatatata tggagttccg cgttacataa cttacggtaa
     3001 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     3061 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     3121 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     3181 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     3241 ctacttggca gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc
     3301 agtacatcaa tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca
     3361 ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta
     3421 acaactccgc cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa
     3481 gcagagctgg tttagtgaac cgtcagatcc gctagcgcta ccggtggcta cccaccatgg
     3541 tgagcaaggg cgaggaggtc atcaaggagt tcatgcgctt caaggagcac atggagggct
     3601 ccgtgaacgg ccacgagttc gagatcgagg gcgagggcga gggccgcccc tacgagggca
     3661 cccagaccgc caggctgaag gtgaccaagg gtggccccct gcccttcgcc tgggacatcc
     3721 tgtcccctca gatcatgtac ggctccaagg cctacgtgaa gcaccccgcc gacatccccg
     3781 actacttgaa gctgtccttc cccgagggct tcaagtggga gcgcgtgatg aacttcgagg
     3841 acggcggcgt ggtgaccgtg acccaggact cctccctgca ggacggcgag ttcatctaca
     3901 aggtgaaggt gcgcggcacc aacttcccct ccgacggccc cgtaatgcag aagaagacca
     3961 tgggctggga ggcctcctcc gagcggatgt accccgagga cggcgccctg aagggcgaga
     4021 tgaagatgag gctgaggctg aaggacggcg gccactacga cgccgaggtc aagaccacct
     4081 acatggccaa gaagcccgtg cagctgcccg gcgcctacaa gaccgacatc aagctggaca
     4141 tcacctccca caacgaggac tacaccatcg tggaacagta cgagcgcgcc gagggccgcc
     4201 actccaccgg cgccttcgaa gataactgat cataatcagc cataccacat ttgtagaggt
     4261 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     4321 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     4381 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     4441 catcaatgta tcttactggg cctcatgggc cgtcgacctc tagctagagc ttggcgtaat
     4501 catggtcatt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4561 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4621 tctggcccca gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca
     4681 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     4741 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     4801 ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     4861 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     4921 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     4981 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5041 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5101 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5161 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5221 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5281 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5341 agggcgacac ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt
     5401 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5461 ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata
     5521 cctgaatatg gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc
     5581 gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5641 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5701 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5761 cgcagatacc aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact
     5821 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5881 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5941 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6001 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     6061 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6121 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6181 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6241 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6301 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6361 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc
     6421 caggcatcaa actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc
     6481 tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa
     6541 cgagtaatcg ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg
     6601 ggagtttagg gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata
     6661 tatgagaatt atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt
     6721 tgctttttcg attgatgaac acctataatt aaactattca tctattattt atgatttttt
     6781 gtatatacaa tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata
     6841 aagggaaaat cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat
     6901 acaaactata agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt
     6961 aattgtgagc ggataacaat tacgagcttc atgcacagtg gcgttgacat tgattattga
     7021 ctagcatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgccatgc
     7081 attagttatt aataacatac gctctccatc aaaacaaaac gaaacaaaac aaactagcaa
     7141 aataggctgt ccccagtgca agtgcaggtg ccagaacatt tctct
//
