LOCUS       Exported                6583 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expresses RFP-HA-DAT in mammalian cells.
ACCESSION   .
VERSION     .
KEYWORDS    RFP-HA-DAT
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6583)
  AUTHORS   Caltagarone J, Ma S, Sorkin A
  TITLE     Dopamine transporter is enriched in filopodia and induces filopodia 
            formation.
  JOURNAL   Mol Cell Neurosci. 2015 Sep;68:120-30. doi: 
            10.1016/j.mcn.2015.04.005. Epub 2015 Apr 30.
  PUBMED    25936602
REFERENCE   2  (bases 1 to 6583)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6583
                     /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"
     regulatory      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1323
                     /codon_start=1
                     /product="monomeric derivative of red fluorescent protein 
                     from Entacmaea quadricolor (Merzlyak et al., 2007)"
                     /label=TagRFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTM
                     RIKVVEGGPLPFAFDILATSFMYGSRTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGG
                     VLTATQDTSLQDGCLIYNVKIRGVNFPSNGPVMQKKTLGWEANTEMLYPADGGLEGRSD
                     MALKLVGGGHLICNFKTTYRSKKPAKNLKMPGVYYVDHRLERIKEADKETYVEQHEVAV
                     ARYCDLPSKLGHKLN"
     protein_bind    1807..1818
                     /label=ZF binding site
                     /bound_moiety="CCR5TC zinc-finger domain"
                     /note="target sequence for the CCR5TC zinc-finger domain 
                     from the CCR5-224 (+) zinc-finger nuclease (Perez et al., 
                     2008; Gross et al., 2013)"
     CDS             1951..1977
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     polyA_signal    3371..3492
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3408..3427)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3462..3481
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3499..3954)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3636..3657)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3848..3867
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3981..4085
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4087..4444
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(4110..4130)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4295..4430
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4357..4376
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4479..5273
                     /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(4533..4552)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5143..5162
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5461..5480)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5505..5552
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5881..6469
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6370..6389
                     /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 gctagcgcta
      601 ccggtcgcca ccatggtgtc taagggcgaa gagctgatta aggagaacat gcacatgaag
      661 ctgtacatgg agggcaccgt gaacaaccac cacttcaagt gcacatccga gggcgaaggc
      721 aagccctacg agggcaccca gaccatgaga atcaaggtgg tcgagggcgg ccctctcccc
      781 ttcgccttcg acatcctggc taccagcttc atgtacggca gcagaacctt catcaaccac
      841 acccagggca tccccgactt ctttaagcag tccttccctg agggcttcac atgggagaga
      901 gtcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccaggac
      961 ggctgcctca tctacaacgt caagatcaga ggggtgaact tcccatccaa cggccctgtg
     1021 atgcagaaga aaacactcgg ctgggaggcc aacaccgaga tgctgtaccc cgctgacggc
     1081 ggcctggaag gcagaagcga catggccctg aagctcgtgg gcgggggcca cctgatctgc
     1141 aacttcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcgtc
     1201 tactatgtgg accacagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     1261 cagcacgagg tggctgtggc cagatactgc gacctcccta gcaaactggg gcacaaactt
     1321 aattccggac tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccatgagt
     1381 aagagcaaat gctccgtggg actcatgtct tccgtggtgg ccccggctaa ggagcccaat
     1441 gccgtgggcc cgaaggaggt ggagctcatc cttgtcaagg agcagaacgg agtgcagctc
     1501 accagctcca ccctcaccaa cccgcggcag agccccgtgg aggcccagga tcgggagacc
     1561 tggggcaaga agatcgactt tctcctgtcc gtcattggct ttgctgtgga cctggccaac
     1621 gtctggcggt tcccctacct gtgctacaaa aatggtggcg gtgccttcct ggtcccctac
     1681 ctgctcttca tggtcattgc tgggatgcca cttttctaca tggagctggc cctcggccag
     1741 ttcaacaggg aaggggccgc tggtgtctgg aagatctgcc ccatactgaa aggtgtgggc
     1801 ttcacggtca tcctcatctc actgtatgtc ggcttcttct acaacgtcat catcgcctgg
     1861 gcgctgcact atctcttctc ctccttcacc acggagctcc cctggatcca ctgcaacaac
     1921 tcctggaaca gccccaactg ctcggatgcc tatccttatg acgtccctga ctacgcctcg
     1981 ctcctcaacg acacttttgg gaccacacct gctgccgagt actttgaacg tggcgtgctg
     2041 cacctccacc agagccatgg catcgacgac ctggggcctc cgcggtggca gctcacagcc
     2101 tgcctggtgc tggtcatcgt gctgctctac ttcagcctct ggaagggcgt gaagacctca
     2161 gggaaggtgg tatggatcac agccaccatg ccatacgtgg tcctcactgc cctgctcctg
     2221 cgtggggtca ccctccctgg agccatagac ggcatcagag catacctgag cgttgacttc
     2281 taccggctct gcgaggcgtc tgtttggatt gacgcggcca cccaggtgtg cttctccctg
     2341 ggcgtggggt tcggggtgct gatcgccttc tccagctaca acaagttcac caacaactgc
     2401 tacagggacg cgattgtcac cacctccatc aactccctga cgagcttctc ctccggcttc
     2461 gtcgtcttct ccttcctggg gtacatggca cagaagcaca gtgtgcccat cggggacgtg
     2521 gccaaggacg ggccagggct gatcttcatc atctacccgg aagccatcgc cacgctccct
     2581 ctgtcctcag cctgggccgt ggtcttcttc atcatgctgc tcaccctggg tatcgacagc
     2641 gccatgggtg gtatggagtc agtgatcacc gggctcatcg atgagttcca gctgctgcac
     2701 agacaccgtg agctcttcac gctcttcatc gtcctagcga ccttcctcct gtccctgttc
     2761 tgcgtcacca acggtggcat ctacgtcttc acgctcctgg accattttgc agccggcacg
     2821 tccatcctct ttggagtgct catcgaagcc atcggagtgg cctggttcta tggtgttggg
     2881 cagttcagcg acgacatcca gcagatgacc gggcagcggc ccagcctgta ctggcggctg
     2941 tgctggaagc tggtcagccc ctgctttctc ctgttcgtgg tcgtggtcag cattgtgacc
     3001 ttcagacccc cccactacgg agcctacatc ttccccgact gggccaacgc gctgggctgg
     3061 gtcatcgcca catcctccat ggccatggtg cccatctatg cggcctacaa gttctgcagc
     3121 ctgcctgggt cctttcgaga gaaactggcc tacgccattg cacccgagaa ggaccgtgag
     3181 ctggtggaca gaggggaggt gcgccagttc acgctccgcc actggctcaa ggtgtagccc
     3241 gggatccacc ggatctagat aactgatcat aatcagccat accacatttg tagaggtttt
     3301 acttgcttta aaaaacctcc cacacctccc cctgaacctg aaacataaaa tgaatgcaat
     3361 tgttgttgtt aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac
     3421 aaatttcaca aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat
     3481 caatgtatct taacgcgtaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     3541 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     3601 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     3661 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     3721 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     3781 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     3841 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     3901 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtcaggtg
     3961 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     4021 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     4081 agagtcctga ggcggaaaga accagctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4141 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     4201 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     4261 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     4321 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     4381 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     4441 caaagatcga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     4501 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     4561 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     4621 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca agacgaggca gcgcggctat
     4681 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4741 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4801 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4861 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4921 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4981 ccgaactgtt cgccaggctc aaggcgagca tgcccgacgg cgaggatctc gtcgtgaccc
     5041 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     5101 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     5161 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     5221 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     5281 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     5341 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     5401 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccctaggg ggaggctaac
     5461 tgaaacacgg aaggagacaa taccggaagg aacccgcgct atgacggcaa taaaaagaca
     5521 gaataaaacg cacggtgttg ggtcgtttgt tcataaacgc ggggttcggt cccagggctg
     5581 gcactctgtc gataccccac cgagacccca ttggggccaa tacgcccgcg tttcttcctt
     5641 ttccccaccc caccccccaa gttcgggtga aggcccaggg ctcgcagcca acgtcggggc
     5701 ggcaggccct gccatagcct caggttactc atatatactt tagattgatt taaaacttca
     5761 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc
     5821 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc
     5881 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
     5941 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
     6001 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
     6061 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
     6121 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
     6181 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
     6241 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
     6301 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
     6361 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
     6421 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
     6481 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
     6541 gttatcccct gattctgtgg ataaccgtat taccgccatg cat
//
