LOCUS       Exported                4861 bp ds-DNA     circular SYN 05-JUL-2022
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4861)
  AUTHORS   Princeton Network
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jul 5, 2022 from SnapGene 5.2.5
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4861
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     misc_feature    98..897
                     /label=EGFR upstream homology
     CDS             910..993
                     /codon_start=1
                     /label=CD3-epsilon
                     /translation="PPPVPNPDYEPIRKGQRDLYSGLNQRRI"
     CDS             1009..1092
                     /codon_start=1
                     /label=CD3-epsilon
                     /translation="PPPVPNPDYEPIRKGQRDLYSGLNQRRI"
     CDS             1108..1191
                     /codon_start=1
                     /label=CD3-epsilon
                     /translation="PPPVPNPDYEPIRKGQRDLYSGLNQRRI"
     CDS             1207..1914
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="VSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYEE
                     LNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFEDG
                     ASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIST
                     FKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQK
                     AFTDVMGMDELYK"
     regulatory      1459..1468
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     misc_feature    1916..2714
                     /label=EGFR downstream homology
     misc_feature    1943..1962
                     /label=EGFR sgRNA2
     misc_feature    1965
                     /label=EDITED
                     /note="G --> C to remove PAM
                     "
     misc_feature    2202
                     /label=EDITED
                     /note="to allow for synthesis by twist (a --> t)
                     "
     primer_bind     2809..2831
                     /label=pGEX 3'
     primer_bind     complement(2869..2887)
                     /label=pBRforEco
     promoter        2955..3059
                     /label=AmpR promoter
     CDS             3060..3920
                     /label=AmpR
     primer_bind     complement(3278..3297)
                     /label=Amp-R
     rep_origin      4091..4679
                     /label=ori
     primer_bind     4580..4599
                     /label=pBR322ori-F
     primer_bind     4833..4850
                     /label=L4440
ORIGIN
        1 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
       61 cgacaggttt cccgactgga aagcgggcag tgagcgcata tacacacacg tacacacaca
      121 ccacacacac ccacatgcac acaccacaca cacatacaaa tatacaccac acacacatac
      181 accacacaca cggtgcacat acacacacat atacacacac cagacacaca taccacatac
      241 acatcacaca tatatgtata catgcataca catacacaca cacatacaca cactctcctc
      301 aaggcagttt atcctctgag aactttaaat ttacaaaaga cacatatgtc cattactttg
      361 agaaggacag gaaagaaccc actttctttt gcagcaacag caagagggcc ctcccgaggc
      421 tcctgctccc tgtcataagt ctccttgttg aggacattca cagggttcag aacccaggga
      481 tcctgcatgg gatggtgctt tgctgattac ttcacctctg atttctttcc actttcagaa
      541 tacataaacc agtccgttcc caaaaggccc gctggctctg tgcagaatcc tgtctatcac
      601 aatcagcctc tgaaccccgc gcccagcaga gacccacact accaggaccc ccacagcact
      661 gcagtgggca accccgagta tctcaacact gtccagccca cctgtgtcaa cagcacattc
      721 gacagccctg cccactgggc ccagaaaggc agccaccaaa ttagcctgga caaccctgac
      781 taccagcagg acttctttcc caaggaagcc aagccaaatg gcatctttaa gggctccaca
      841 gctgaaaatg cagaatacct aagggtcgcg ccacaaagca gtgaatttat tggagcaggc
      901 tctggttctc cacctcccgt ccctaatccc gattatgagc ccattcggaa aggtcaaaga
      961 gatttgtata gtggcctcaa tcaaagaaga atcgggtcag gttccggccc gccgccagtc
     1021 ccaaaccccg actatgaacc aatcaggaaa ggccaacggg atttgtactc aggattgaat
     1081 cagcgcagga taggttccgg ctccgggcca ccacccgtcc ctaacccaga ttacgaaccc
     1141 atacgcaagg gacaacgcga tctttacagc ggattgaacc aacgacgaat aggatctggt
     1201 tctggcgtga gcaagggcga ggaggataac atggcctctc tcccagcgac acatgagtta
     1261 cacatctttg gctccatcaa cggtgtggac tttgacatgg tgggtcaggg caccggcaat
     1321 ccaaatgatg gttatgagga gttaaacctg aagtccacca agggtgacct ccagttctcc
     1381 ccctggattc tggtccctca tatcgggtat ggcttccatc agtacctgcc ctaccctgac
     1441 gggatgtcgc ctttccaggc cgccatggta gatggctccg gataccaagt ccatcgcaca
     1501 atgcagtttg aagatggtgc ctcccttact gttaactacc gctacaccta cgagggaagc
     1561 cacatcaaag gagaggccca ggtgaagggg actggtttcc ctgctgacgg tcctgtgatg
     1621 accaactcgc tgaccgctgc ggactggtgc aggtcgaaga agacttaccc caacgacaaa
     1681 accatcatca gtacctttaa gtggagttac accactggaa atggcaagcg ctaccggagc
     1741 actgcgcgga ccacctacac ctttgccaag ccaatggcgg ctaactatct gaagaaccag
     1801 ccgatgtacg tgttccgtaa gacggagctc aagcactcca agaccgagct caacttcaag
     1861 gagtggcaaa aggcctttac cgatgtgatg ggcatggacg agctgtacaa gtaaccacgg
     1921 aggatagtat gagccctaaa aatccagact ctttcgatac ccagcaccaa gccacagcag
     1981 gtcctccatc ccaacagcca tgcccgcatt agctcttaga cccacagact ggttttgcaa
     2041 cgtttacacc gactagccag gaagtacttc cacctcgggc acattttggg aagttgcatt
     2101 cctttgtctt caaactgtga agcatttaca gaaacgcatc cagcaagaat attgtccctt
     2161 tgagcagaaa tttatctttc aaagaggtat atttgaaaaa ataaaaaagt atatgtgagg
     2221 atttttattg attggggatc ttggagtttt tcattgtcgc tattgatttt tacttcaatg
     2281 ggctcttcca acaaggaaga agcttgctgg tagcacttgc taccctgagt tcatccaggc
     2341 ccaactgtga gcaaggagca caagccacaa gtcttccaga ggatgcttga ttccagtggt
     2401 tctgcttcaa ggcttccact gcaaaacact aaagatccaa gaaggccttc atggccccag
     2461 caggccggat cggtactgta tcaagtcatg gcaggtacag taggataagc cactctgtcc
     2521 cttcctgggc aaagaagaaa cggaggggat ggaattcttc cttagactta cttttgtaaa
     2581 aatgtcccca cggtacttac tccccactga tggaccagtg gtttccagtc atgagcgtta
     2641 gactgacttg tttgtcttcc attccattgt tttgaaactc agtatgctgc ccctgtcttg
     2701 ctgtcatgaa atcattaagc cagccccgac acccgccaac acccgctgac gcgccctgac
     2761 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     2821 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac
     2881 gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt
     2941 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     3001 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     3061 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     3121 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     3181 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     3241 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     3301 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     3361 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     3421 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     3481 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     3541 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     3601 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     3661 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     3721 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     3781 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     3841 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     3901 cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt
     3961 taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga
     4021 ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca
     4081 aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac
     4141 caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg
     4201 taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag
     4261 gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac
     4321 cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt
     4381 taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg
     4441 agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc
     4501 ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc
     4561 gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc
     4621 acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa
     4681 acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt
     4741 tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg
     4801 ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag
     4861 a
//
