LOCUS       Exported                5564 bp ds-DNA     circular SYN 04-NOV-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5564)
  AUTHORS   Jiten Doshi
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Nov 4, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5564
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    45..88
                     /label=T7
                     /label=nonstandard type: misc_feature_
     terminator      complement(46..92)
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     polyA_signal    110..158
                     /note="synthetic polyadenylation signal"
     primer_bind     189..205
                     /label=PR-JD21
                     /note="fp to amplify ef1a promoter from pJD02 to transfer 
                     to pJD10; Gibson assembly"
     misc_feature    190..1373
                     /label=EF1a
                     /label=nonstandard type: misc_feature_
     intron          426..1364
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1407..1426
                     /label=PR-JD47
                     /note="sequencing primer pJD01"
     primer_bind     complement(1449..1469)
                     /label=PR-JD22
                     /note="rp to amplify ef1a promoter from pJD02 to transfer 
                     to pJD10; Gibson assembly"
     misc_feature    1470..1583
                     /label=insert
                     /label=nonstandard type: misc_feature_
     misc_feature    1470..1569
                     /label=synthetic 5' UTR
                     /note="/from pJD48; pRA16"
     primer_bind     1470..1493
                     /label=PR-JD58
                     /note="syn5utr reverse primer for part2 golden gate 
                     verification; (150bp product)"
     primer_bind     1564..1603
                     /label=Fragment.FOR
     misc_feature    1579..1583
                     /gene="Kozak sequence"
                     /label=Kozak sequence
     regulatory      1579..1583
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1585..2199
                     /codon_start=1
                     /label=23K endoprotease protease
                     /note="vntifkey=21"
                     /translation="MGSSEQELKAIVKDLGCGPYFLGTYDKRFPGFVSPHKLACAIVNT
                     AGRETGGVHWMAFAWNPHSKTCYLFEPFGFSDQRLKQVYQFEYESLLRRSAIASSPDRC
                     ITLEKSTQSVQGPNSAACGLFCCMFLHAFANWPQTPMDHNPTMNLITGVPNSMLNSPQV
                     QPTLRRNQEQLYSFLERHSPYFRSHSAQIRSATSFCHLKNM"
     primer_bind     complement(2167..2224)
                     /label=Fragment.REV
     misc_feature    join(2205..5564,1..1469)
                     /label=vector_backbone-835-5669
                     /label=nonstandard type: misc_feature_
     misc_feature    2206..2232
                     /label=junk bacterial sequence??
                     /note="blast shows match to bacterial sequences!"
     misc_feature    2233..2466
                     /label=SV40_PA
                     /label=nonstandard type: misc_feature_
     primer_bind     complement(2233..2252)
                     /label=PR-JD48
                     /note="sequencing primer pJD01"
     misc_feature    2305..2310
                     /label=pA signal (bottom strand)
     primer_bind     2314..2333
                     /label=SV40pA-R
                     /note="Genewiz primer collection"
     misc_feature    2334..2339
                     /label=pA signal (bottom strand)
     misc_feature    2359..2364
                     /label=pA signal
     primer_bind     2404..2420
                     /label=EGFP-C-REV
                     /note="Genewiz primer collection"
     terminator      2485..2531
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     terminator      2561..2697
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    2742..2813
                     /label=3' beta-globin insulator
                     /note="insulator downstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
     misc_feature    3039..3627
                     /label=colE1_high_copy
                     /label=nonstandard type: misc_feature_
     rep_origin      complement(3039..3627)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3084..3103)
                     /label=PR-JD29
                     /note="pJD11 rtTA CDS, pA sequencing primer"
     primer_bind     3645..3663
                     /label=PR-JD32
                     /note="pJD02/03 sequencing primer"
     misc_feature    4373..5233
                     /label=ampR
                     /label=nonstandard type: misc_feature_
     CDS             4373..5233
                     /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"
     terminator      5246..5382
                     /label=rrnG terminator
                     /note="transcription terminator from the E. coli ribosomal 
                     RNA rrnG operon (Albrechtsen et al., 1991)"
     misc_feature    join(5534..5564,1..41)
                     /label=5' beta-globin insulator
                     /note="insulator upstream of the human beta-globin locus 
                     (Farrell et al., 2002)"
ORIGIN
        1 cttccctcta gtggtcataa cagcagcttc agctacctct caattcaaaa aacccctcaa
       61 gacccgttta gaggccccaa ggggttatgc tatcaatcgt tgcgttacac acacaaaaaa
      121 ccaacacaca tccatcttcg atggatagcg attttattat ctaactgctg atcgagtgta
      181 gccagatctc gtgaggctcc ggtgcccgtc agtgggcaga gcgcacatcg cccacagtcc
      241 ccgagaagtt ggggggaggg gtcggcaatt gaaccggtgc ctagagaagg tggcgcgggg
      301 taaactggga aagtgatgtc gtgtactggc tccgcctttt tcccgagggt gggggagaac
      361 cgtatataag tgcactagtc gccgtgaacg ttctttttcg caacgggttt gccgccagaa
      421 cacaggtaag tgccgtgtgt ggttcccgcg ggcctggcct ctttacgggt tatggccctt
      481 gcgtgccttg aattacttcc acctggctgc agtacgtgat tcttgatccc gagcttcggg
      541 ttggaagtgg gtgggagagt tcgtggcctt gcgcttaagg agccccttcg cctcgtgctt
      601 gagttgtggc ctggcctggg cgctggggcc gccgcgtgcg aatctggtgg caccttcgcg
      661 cctgtctcgc tgctttcgat aagtctctag ccatttaaaa tttttgatga cctgctgcga
      721 cgcttttttt ctggcaagat agtcttgtaa atgcgggcca agatcagcac actggtattt
      781 cggtttttgg ggccgcgggc ggcgacgggg cccgtgcgtc ccagcgcaca tgttcggcga
      841 ggcggggcct gcgagcgcgg ccaccgagaa tcggacgggg gtagtctcaa gctgcccggc
      901 ctgctctggt gcctggcctc gcgccgccgt gtatcgcccc gccctgggcg gcaaggctgg
      961 cccggtcggc accagttgcg tgagcggaaa gatggccgct tcccggccct gctgcaggga
     1021 gcacaaaatg gaggacgcgg cgctcgggag agcgggcggg tgagtcaccc acacaaagga
     1081 aaagggcctt tccgtcctca gccgtcgctt catgtgactc cacggagtac cgggcgccgt
     1141 ccaggcacct cgattagttc tccagctttt ggagtacgtc gtctttaggt tggggggagg
     1201 ggttttatgc gatggagttt ccccacactg agtgggtgga gactgaagtt aggccagctt
     1261 ggcacttgat gtaattctcc ttggaatttg ccctttttga gtttggatct tggttcattc
     1321 tcaagcctca gacagtggtt caaagttttt ttcttccatt tcaggtgtcg tgaaaactac
     1381 ccctaaaagc caaaagatct ttgtcgatcc taccatccac tcgacacacc cgccagcggc
     1441 cgctgccaag cttccgagct ctcgaattca tcttggctga ggaatcttct aacaatttag
     1501 agcttaaaaa cgcccacgag gcggagaacg aaatatccag agagacgtta gaaacgttca
     1561 aaaacgttca ccggtagggc caccatgggc tccagtgagc aggaactgaa agccattgtc
     1621 aaagatcttg gttgtgggcc atattttttg ggcacctatg acaagcgctt tccaggcttt
     1681 gtttctccac acaagctcgc ctgcgccata gtcaatacgg ccggtcgcga gactgggggc
     1741 gtacactgga tggcctttgc ctggaacccg cactcaaaaa catgctacct ctttgagccc
     1801 tttggctttt ctgaccagcg actcaagcag gtttaccagt ttgagtacga gtcactcctg
     1861 cgccgtagcg ccattgcttc ttcccccgac cgctgtataa cgctggaaaa gtccacccaa
     1921 agcgtacagg ggcccaactc ggccgcctgt ggactattct gctgcatgtt tctccacgcc
     1981 tttgccaact ggccccaaac tcccatggat cacaacccca ccatgaacct tattaccggg
     2041 gtacccaact ccatgctcaa cagtccccag gtacagccca ccctgcgtcg caaccaggaa
     2101 cagctctaca gcttcctgga gcgccactcg ccctacttcc gcagccacag tgcgcagatt
     2161 aggagcgcca cttctttttg tcacttgaaa aacatgtaag ctagcttgac tgactgagat
     2221 acagcgtacc ttcagctcac agacatgata agatacattg atgagtttgg acaaaccaca
     2281 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     2341 gtaaccatta taagctgcaa taaacaagtt aacaacaaca attgcattca ttttatgttt
     2401 caggttcagg gggaggtgtg ggaggttttt taaagcaagt aaaacctcta caaatgtggt
     2461 attggcccat ctctatcggt atcgtagcat aaccccttgg ggcctctaaa cgggtcttga
     2521 ggggtttttt gtgcccctcg ggccggattg ctatctaccg gcattggcgc agaaaaaaat
     2581 gcctgatgcg acgctgcgcg tcttatactc ccacatatgc cagattcagc aacggatacg
     2641 gcttccccaa cttgcccact tccatacgtg tcctccttac cagaaattta tccttaaggt
     2701 cgtcagctat cctgcaggcg atctctcgat ttcgatcaag acattccttt aatggtcttt
     2761 tctggacacc actaggggtc agaagtagtt catcaaactt tcttccctcc ctaatctcat
     2821 tggttacctt gggctatcga aacttaatta accagtcaag tcagctactt ggcgagatcg
     2881 acttgtctgg gtttcgacta cgctcagaat tgcgtcagtc aagttcgatc tggtccttgc
     2941 tattgcaccc gttctccgat tacgagtttc atttaaatca tgtgagcaaa aggccagcaa
     3001 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     3061 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     3121 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     3181 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3241 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3301 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3361 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     3421 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     3481 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     3541 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     3601 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     3661 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     3721 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     3781 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     3841 ctatttcgtt catccatagt tgcatttaaa tttccgaact ctccaaggcc ctcgtcggaa
     3901 aatcttcaaa cctttcgtcc gatccatctt gcaggctacc tctcgaacga actatcgcaa
     3961 gtctcttggc cggccttgcg ccttggctat tgcttggcag cgcctatcgc caggtattac
     4021 tccaatcccg aatatccgag atcgggatca cccgagagaa gttcaaccta catcctcaat
     4081 cccgatctat ccgagatccg aggaatatcg aaatcggggc gcgcctggtg taccgagaac
     4141 gatcctctca gtgcgagtct cgacgatcca tatcgttgct tggcagtcag ccagtcggaa
     4201 tccagcttgg gacccaggaa gtccaatcgt cagatattgt actcaagcct ggtcacggca
     4261 gcgtaccgat ctgtttaaac ctagatattg atagtctgat cggtcaacgt ataatcgagt
     4321 cctagctttt gcaaacatct atcaagagac aggatcagca ggaggctttc gcatgagtat
     4381 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     4441 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cgcgagtggg
     4501 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     4561 ctttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     4621 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     4681 ttcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     4741 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga ttggaggacc
     4801 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     4861 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     4921 aatggcaaca accttgcgta aactattaac tggcgaacta cttactctag cttcccggca
     4981 acagttgata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     5041 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     5101 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     5161 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     5221 taagcattgg taaccgattc taggtgcatt ggcgcagaaa aaaatgcctg atgcgacgct
     5281 gcgcgtctta tactcccaca tatgccagat tcagcaacgg atacggcttc cccaacttgc
     5341 ccacttccat acgtgtcctc cttaccagaa atttatcctt aagatcccga atcgtttaaa
     5401 ctcgactctg gctctatcga atctccgtcg tttcgagctt acgcgaacag ccgtggcgct
     5461 catttgctcg tcgggcatcg aatctcgtca gctatcgtca gcttaccttt ttggcagcga
     5521 tcgcggctcc cgacatcttg gaccattagc tccacaggta tctt
//
