LOCUS       Exported                3440 bp ds-DNA     circular SYN 25-JAN-2023
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3440)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Apr 5, 2023 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..3440
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     terminator      1..87
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     21..40
                     /label=A423 rrnb T1 ter -f
                     /note="color: black; sequence: cgttcaccgacaaacaacag; added:
                     2022-07-25"
     primer_bind     127..146
                     /label=C133 pME 5' con -f
                     /note="color: black; sequence: ggatgttttcccagtcacga; added:
                     2022-07-25"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     182..219
                     /label=kozak-EGFP-BsaI-AGTC-f
                     /note="color: black; sequence: 
                     atcggtctctAGTCgccaccatggtgagcaagggcgagga; added: 
                     2022-08-02"
     primer_bind     194..237
                     /label=A835 vec new-f ryan
                     /note="color: black; sequence: 
                     TCCTGAAGCCACCatggtgagcaagggcgaggagctgttcaccggggtggt; added:
                     2023-01-25"
     regulatory      194..203
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             200..916
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="This feature has 3 segments:
                        1: 200 .. 202 / #05fc14
                        2: 203 .. 205 / #05fc14 / 1a
                        3: 206 .. 916 / #05fc14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(200..220)
                     /label=YFP start -r
                     /note="color: black; sequence: CTCCTCGCCCTTGCTCACCAT; 
                     added: 2023-01-25"
     primer_bind     complement(245..266)
                     /label=A228 EGFP-N
                     /note="color: black; sequence: cgtcgccgtccagctcgaccag; 
                     added: 2022-02-04"
     primer_bind     complement(267..286)
                     /label=C151 gfp con 2nd -r
                     /note="color: black; sequence: gctgaacttgtggccgttta; added:
                     2023-01-25"
     primer_bind     complement(678..698)
                     /label=C19 GFP confirm -r
                     /note="color: black; sequence: tgaagttcaccttgatgccgt; 
                     added: 2022-02-04"
     primer_bind     840..859
                     /label=C18 GFP confirm -f
                     /note="color: black; sequence: agaagcgcgatcacatggtc; added:
                     2023-01-25"
     primer_bind     complement(899..919)
                     /label=C17 mCherry stop XhoI -r
                     /note="color: black; sequence: 
                     cctcgagctacttgtacagctcgtccat; added: 2023-01-25"
     promoter        complement(926..944)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    949..1083
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     962..989
                     /label=C251 FRT-a-cry-Ds-Red-SV40pA-FRT -r
                     /note="color: black; sequence: 
                     ggcggatccgaagttcctattctctagaaagtataggaacttctaagatacattgatga
                      gtttggacaaac; added: 2023-01-25"
     primer_bind     complement(1127..1160)
                     /label=SV40-pA-BsaI-CTCA-r
                     /note="color: black; sequence: 
                     gatggtctctTGAGgaattaaaaaacctcccacacct; added: 2022-08-02"
     CDS             1520..2311
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /note="color: #ccffcc"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      2404..2992
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     2893..2912
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     3146..3163
                     /label=A757 L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     terminator      3322..3349
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      121 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      181 ccggtctcta gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc
      241 catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg
      301 cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca ccggcaagct
      361 gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg
      421 ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg aaggctacgt
      481 ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg ccgaggtgaa
      541 gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact tcaaggagga
      601 cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg tctatatcat
      661 ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca acatcgagga
      721 cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg acggccccgt
      781 gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag accccaacga
      841 gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca ctctcggcat
      901 ggacgagctg tacaagtagt ctagaactat agtgagtcgt attacgtaga tccagacatg
      961 ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa aaaatgcttt
     1021 atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg caataaacaa
     1081 gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt gtgggaggtt
     1141 ttttaattcc tcaagagacc tctagaccag ccaggacaga aatgcctcga cttcgctgct
     1201 acccaaggtt gccgggtgac gcacaccgtg gaaacggatg aaggcacgaa cccagtggac
     1261 ataagcctgt tcggttcgta agctgtaatg caagtagcgt atgcgctcac gcaactggtc
     1321 cagaaccttg accgaacgca gcggtggtaa cggcgcagtg gcggttttca tggcttgtta
     1381 tgactgtttt tttggggtac agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc
     1441 cgtgggtcga tgtttgatgt tatggagcag caacgatgtt acgcagcagg gcagtcgccc
     1501 taaaacaaag ttaaacatta tgagggaagc ggtgatcgcc gaagtatcga ctcaactatc
     1561 agaggtagtt ggcgtcatcg agcgccatct cgaaccgacg ttgctggccg tacatttgta
     1621 cggctccgca gtggatggcg gcctgaagcc acacagtgat attgatttgc tggttacggt
     1681 gaccgtaagg cttgatgaaa caacgcggcg agctttgatc aacgaccttt tggaaacttc
     1741 ggcttcccct ggagagagcg agattctccg cgctgtagaa gtcaccattg ttgtgcacga
     1801 cgacatcatt ccgtggcgtt atccagctaa gcgcgaactg caatttggag aatggcagcg
     1861 caatgacatt cttgcaggta tcttcgagcc agccacgatc gacattgatc tggctatctt
     1921 gctgacaaaa gcaagagaac atagcgttgc cttggtaggt ccagcggcgg aggaactctt
     1981 tgatccggtt cctgaacagg atctatttga ggcgctaaat gaaaccttaa cgctatggaa
     2041 ctcgccgccc gactgggctg gcgatgagcg aaatgtagtg cttacgttgt cccgcatttg
     2101 gtacagcgca gtaaccggca aaatcgcgcc gaaggatgtc gctgccgact gggcaatgga
     2161 gcgcctgccg gcccagtatc agcccgtcat acttgaagct agacaggctt atcttggaca
     2221 agaagaagat cgcttggcct cgcgcgcaga tcagttggaa gaatttgtcc actacgtgaa
     2281 aggcgagatc accaaggtag tcggcaaata accctcgagc cacccatgac caaaatccct
     2341 taacgtgagt tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2401 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2461 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2521 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2581 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2641 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2701 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2761 gacctacacc gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga
     2821 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     2881 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     2941 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3001 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3061 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3121 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     3181 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     3241 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta ccgctagcca
     3301 ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg cttagtttga
     3361 tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg cttcacaacg
     3421 ttcaaatccg ctcccggcgg
//
