LOCUS       Exported                5193 bp ds-DNA     circular SYN 04-APR-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5193)
  AUTHORS   Jonathan Pacheco
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, May 18, 2023 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5193
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     565..589
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     regulatory      601..610
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             607..1314
                     /codon_start=1
                     /product="photoactivatable monomeric derivative of DsRed 
                     fluorescent protein (Subach et al., 2009)"
                     /label=PAmCherry1
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHVFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFTWDILSPQFMYGSNAYVKHPADIPDYFKLSFPEGFKWERVMKF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEALSERMYPEDGALK
                     GEVKPRVKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNRKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(752..770)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1020..1039
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     misc_feature    1315..1337
                     /label=MCS
                     /note="multiple cloning site"
     CDS             1354..1851
                     /codon_start=1
                     /gene="sidC"
                     /locus_tag="lpg2511"
                     /product="protein SidC, interaptin"
                     /function="Toxin production / other pathogen functions"
                     /label=sidC
                     /db_xref="GeneID:19834076"
                     /protein_id="YP_096518.1"
                     /translation="SKYSSKPLLDVELNKIAEGLELTAKIYNEKRGREWWFKGSRNEAR
                     KTQCEELQRVSKEINTLLQSESLTKSQVLEKVLNSIETLDKIDRDISAESNWFQSTLQK
                     EVRLFRDQLKDICQLDKYAFKSTKLDEIISLEMEEQFQKIQDPAVQQIVRDLPSHCHND
                     EAI"
     gene            1354..1851
                     /gene="sidC"
                     /locus_tag="lpg2511"
                     /label=sidC
                     /db_xref="GeneID:19834076"
     misc_feature    1853..1857
                     /label=MCS
                     /note="multiple cloning site"
     polyA_signal    1981..2102
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2018..2037)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2072..2091
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2109..2564)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2246..2267)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2458..2477
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2591..2695
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2697..3054
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(2720..2740)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2905..3040
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2967..2986
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(2990..3001)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3089..3883
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3143..3162)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3753..3772
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4071..4090)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4115..4162
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4491..5079
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4980..4999
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcggtc
      601 gccaccatgg tgagcaaggg cgaggaggat aacatggcca tcattaagga gttcatgcgc
      661 ttcaaggtgc acatggaggg gtccgtgaac ggccacgtgt tcgagatcga gggcgagggc
      721 gagggccgcc cctacgaggg cacccagacc gccaagctga aggtgaccaa gggcggcccc
      781 ctgcccttca cctgggacat cctgagccct cagttcatgt acggctccaa tgcctacgtg
      841 aagcaccccg ccgacatccc cgactacttt aagctgtcct tccccgaggg cttcaagtgg
      901 gagcgcgtga tgaaattcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg
      961 caggacggcg agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc
     1021 cccgtgatgc agaagaagac catgggctgg gaggccctct ccgagcggat gtaccccgag
     1081 gacggcgccc tgaagggcga ggtcaagccc agagtgaagc tgaaggacgg cggccactac
     1141 gacgctgagg tcaagaccac ctacaaggcc aagaagcccg tgcagctgcc cggcgcctac
     1201 aacgtcaacc gcaagctgga catcaccagc cacaacgagg actacaccat cgtggagcag
     1261 tacgagagag ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtccgga
     1321 ctcagatctc gagctcaagc ttcgaattca atgtcaaaat attcctccaa gccattattg
     1381 gatgtggaat taaataaaat cgctgaagga ttagaactca ctgcaaaaat ttacaatgaa
     1441 aagagaggac gcgaatggtg gtttaaaggt tcaagaaatg aagctcgtaa gacccaatgt
     1501 gaagaattgc aaagagtatc caaagaaatc aatactcttc tgcaaagtga atctttaacg
     1561 aaaagccagg tacttgaaaa ggttttaaat tctatagaaa cattagataa aattgacaga
     1621 gacatttctg ccgaatccaa ttggtttcaa agtactctgc aaaaggaagt caggttattt
     1681 cgagatcaat tgaaagatat ttgccaattg gacaagtatg cctttaaatc aacaaaactt
     1741 gatgaaatca tctctctgga aatggaagaa caatttcaaa agatacaaga tcctgctgtt
     1801 caacaaattg tcagggactt gccttctcat tgccacaatg atgaagcaat tggatccacc
     1861 ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt acttgcttta
     1921 aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt
     1981 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca
     2041 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct
     2101 taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     2161 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     2221 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     2281 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     2341 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     2401 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     2461 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     2521 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg gcacttttcg
     2581 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     2641 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtcctga
     2701 ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc
     2761 ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag
     2821 tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc
     2881 atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct
     2941 ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc ctcggcctct
     3001 gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg caaagatcga
     3061 tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc acgcaggttc
     3121 tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga caatcggctg
     3181 ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt ttgtcaagac
     3241 cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat cgtggctggc
     3301 cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg gaagggactg
     3361 gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg ctcctgccga
     3421 gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc cggctacctg
     3481 cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga tggaagccgg
     3541 tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag ccgaactgtt
     3601 cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc atggcgatgc
     3661 ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg actgtggccg
     3721 gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata ttgctgaaga
     3781 gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg ctcccgattc
     3841 gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac tctggggttc
     3901 gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc caccgccgcc
     3961 ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat gatcctccag
     4021 cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac tgaaacacgg
     4081 aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca gaataaaacg
     4141 cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc
     4201 gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc
     4261 caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct
     4321 gccatagcct caggttactc atatatactt tagattgatt taaaacttca tttttaattt
     4381 aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc ttaacgtgag
     4441 ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     4501 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     4561 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     4621 cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt caagaactct
     4681 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     4741 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     4801 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     4861 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     4921 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     4981 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     5041 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     5101 ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc gttatcccct
     5161 gattctgtgg ataaccgtat taccgccatg cat
//
