LOCUS       Exported                4771 bp ds-DNA     circular SYN 22-APR-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4771)
  AUTHORS   Jonathan Pacheco
  TITLE     Direct Submission
  JOURNAL   Exported Friday, May 19, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4771
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..155
                     /label=CMV
                     /note="CMV"
     misc_feature    1..155
                     /label=CMVd
     misc_feature    204..713
                     /label=PH-PLCD1
                     /note="PLCD_GFP"
     primer_bind     complement(693..740)
                     /label=PHPLC-NGxR
     misc_feature    726..1433
                     /label=mNG
     CDS             726..1433
                     /codon_start=1
                     /product="bright monomeric yellow-green fluorescent protein
                     derived from LanYFP (Shaner et al., 2013)"
                     /label=mNeonGreen
                     /translation="MVSKGEEDNMASLPATHELHIFGSINGVDFDMVGQGTGNPNDGYE
                     ELNLKSTKGDLQFSPWILVPHIGYGFHQYLPYPDGMSPFQAAMVDGSGYQVHRTMQFED
                     GASLTVNYRYTYEGSHIKGEAQVKGTGFPADGPVMTNSLTAADWCRSKKTYPNDKTIIS
                     TFKWSYTTGNGKRYRSTARTTYTFAKPMAANYLKNQPMYVFRKTELKHSKTELNFKEWQ
                     KAFTDVMGMDELYK"
     CDS             726..746
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     CDS             726..746
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MVSKGEE"
     misc_feature    726..746
                     /label=EGFP
                     /note="EGFP"
     primer_bind     726..743
                     /label=FPi-F
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     726..743
                     /label=NGx-PHPLC-F
     misc_feature    726..740
                     /label=mCherry
                     /note="pmCherry-N1"
     primer_bind     729..746
                     /label=FPi-F2.0
     primer_bind     complement(729..746)
                     /label=FPv-R
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     regulatory      981..990
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     primer_bind     1315..1335
                     /label=NeonGFWD1
     CDS             1414..1433
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     CDS             1414..1433
                     /codon_start=1
                     /label=EGFP
                     /note="EGFP"
                     /translation="MDELYK"
     misc_feature    1414..1433
                     /label=EGFP
                     /note="EGFP"
     primer_bind     complement(1414..1433)
                     /label=FPi-R
                     /note="Amplify fluorescent protein from eGFP/CFP etc pr 
                     mCherry etc"
     primer_bind     1414..1433
                     /label=FPv-F
                     /note="Amplify vector without eGFP/CFP etc pr mCherry etc. 
                     Use to exchange with FPi primers."
     primer_bind     complement(1465..1484)
                     /label=SV40pA-R
     misc_feature    1687..2142
                     /label=f1
                     /note="f1"
     misc_feature    1714..2142
                     /label=f1
                     /note="f1"
     misc_feature    complement(1729..2142)
                     /label=f1ori
                     /note="f1"
     misc_feature    1819..2125
                     /label=f1( )
                     /note="f1( )"
     promoter        2204..2232
                     /label=Bacterial
                     /note="Bacterial"
     promoter        2316..2545
                     /label=SV40early
                     /note="SV40early"
     misc_feature    2483..2618
                     /label=SV40
                     /note="SV40"
     misc_feature    2667..3461
                     /label=Kan/Neo
                     /note="Kan/Neo"
     misc_feature    3697..3715
                     /label=HSV-TK
                     /note="HSV-TK"
     misc_feature    4046..4689
                     /label=pUC
                     /note="pUC"
     primer_bind     complement(4752..4771)
                     /label=NGx-PHPLC-R
     primer_bind     join(4755..4771,1..21)
                     /label=PHPLC-NGxF
ORIGIN
        1 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
       61 gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      121 atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg actcagatct
      181 cgagctcaag cttcgaattc ggcatggact cgggccggga cttcctgacc ctgcacggcc
      241 tacaggatga tgaggatcta caggcgctgc tgaagggcag ccagctcctg aaggtgaagt
      301 ccagctcatg gaggagagag cggttctaca agttgcagga ggactgcaag accatctggc
      361 aggagtcccg caaggtcatg cggaccccgg agtcccagct gttctccatc gaggacattc
      421 aggaggtgcg aatggggcac cgcacggagg gtctggagaa gttcgcccgt gatgtgcccg
      481 aggaccgctg cttctccatt gtcttcaagg accagcgcaa tacactagac ctcatcgccc
      541 catcgccagc tgatgcccag cactgggtgc tggggctgca caagatcatc caccactcag
      601 gctccatgga ccagcgtcag aagctacagc actggattca ctcctgcttg cgaaaagctg
      661 acaaaaacaa ggacaacaag atgagcttca aggagctgca gaacttcctg aagggggtgg
      721 gtggcatggt gagcaagggc gaggaggata acatggcctc tctcccagcg acacatgagt
      781 tacacatctt tggctccatc aacggtgtgg actttgacat ggtgggtcag ggcaccggca
      841 atccaaatga tggttatgag gagttaaacc tgaagtccac caagggtgac ctccagttct
      901 ccccctggat tctggtccct catatcgggt atggcttcca tcagtacctg ccctaccctg
      961 acgggatgtc gcctttccag gccgccatgg tagatggctc cggataccaa gtccatcgca
     1021 caatgcagtt tgaagatggt gcctccctta ctgttaacta ccgctacacc tacgagggaa
     1081 gccacatcaa aggagaggcc caggtgaagg ggactggttt ccctgctgac ggtcctgtga
     1141 tgaccaactc gctgaccgct gcggactggt gcaggtcgaa gaagacttac cccaacgaca
     1201 aaaccatcat cagtaccttt aagtggagtt acaccactgg aaatggcaag cgctaccgga
     1261 gcactgcgcg gaccacctac acctttgcca agccaatggc ggctaactat ctgaagaacc
     1321 agccgatgta cgtgttccgt aagacggagc tcaagcactc caagaccgag ctcaacttca
     1381 aggagtggca aaaggccttt accgatgtga tgggcatgga cgagctgtac aagtaaagcg
     1441 gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac ttgctttaaa
     1501 aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg ttgttgttaa
     1561 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     1621 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     1681 aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca
     1741 gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga
     1801 ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg
     1861 actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat
     1921 caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag
     1981 ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga
     2041 agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa
     2101 ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc acttttcggg
     2161 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     2221 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtcctgagg
     2281 cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     2341 agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt gtggaaagtc
     2401 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccat
     2461 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2521 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2581 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aagatcgatc
     2641 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     2701 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     2761 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     2821 acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg tggctggcca
     2881 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     2941 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     3001 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     3061 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     3121 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     3181 ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     3241 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     3301 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     3361 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     3421 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga
     3481 aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt
     3541 ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg
     3601 cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg aaacacggaa
     3661 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3721 cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc actctgtcga
     3781 taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt ccccacccca
     3841 ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg caggccctgc
     3901 catagcctca ggttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     3961 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     4021 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     4081 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     4141 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     4201 gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     4261 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     4321 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     4381 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     4441 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     4501 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     4561 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     4621 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     4681 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     4741 ttctgtggat aaccgtatta ccgccatgca t
//
