LOCUS       sequence-44486-e        5858 bp    DNA     circular SYN 14-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5858)
  AUTHORS   Mousseau G, Clementz MA, Bakeman WN, Nagarsheth N, Cameron M, Shi J,
            Baran P, Fromentin R, Chomont N, Valente ST
  TITLE     An Analog of the Natural Steroidal Alkaloid Cortistatin A Potently 
            Suppresses Tat-Dependent HIV Transcription.
  JOURNAL   Cell Host Microbe. 2012 Jul 19;12(1):97-108.
  PUBMED    22817991
REFERENCE   2  (bases 1 to 5858)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell Host 
            Microbe."; date: "2012-07-19"; volume: "12(1)"; pages: "97-108"
COMMENT     Sequence Label: pcDNA3.1-TAT-2-130-FLAG
FEATURES             Location/Qualifiers
     source          1..5858
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(44..63)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        235..614
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        615..818
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     769..789
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     863..882
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        863..881
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             1301..1324
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     CDS             1356..1397
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     CDS             1407..1424
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1447..1464)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1453..1677
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1723..2151
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1810..1829)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2020..2041
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2160..2180)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2165..2494
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2345..2480
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2407..2426
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2561..3355
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2615..2634)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3225..3244
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3531..3652
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3568..3587)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3622..3641
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3701..3717)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3701..3717)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3714..3736)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3725..3741
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(3749..3755,3756..3773,3774..3779))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3794..3815
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3932..3949)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4103..4691)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4183..4202)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4862..5653,5654..5722))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5485..5504
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5723..5827)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagt
      901 taagcttacc atggagacac ccttgaaggc gccagagagc tcattaaagt cctgcaacga
      961 gcccttttca cgcacttcag agcaggatgt ggccactcaa gaattggcca gacaagggga
     1021 ggaaatcctc tctcagctat accgacccct agaaacatgc aataactcat gctattgtaa
     1081 gcgatgctgc taccattgtc agatgtgttt tctaaacaag gggctcggga tatgttatga
     1141 acgaaagggc agacgaagaa ggactccaaa gaaaactaag actcatccgt ctcctacacc
     1201 agacaaatcg atatccacaa ggaccgggga cagccagcca acgaagaaac agaagaagac
     1261 ggtggaagca acggtggaga cagatactgg ccctggccga gactacaagg acgacgatga
     1321 caagtagacg cagtctagag ggcccgcggt tcgaaggtaa gcctatccct aaccctctcc
     1381 tcggtctcga ttctacgcgt accggtcatc atcaccatca ccattgagtt taaacccgct
     1441 gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc tcccccgtgc
     1501 cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat gaggaaattg
     1561 catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg caggacagca
     1621 agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc tctatggctt
     1681 ctgaggcgga aagaaccagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     1741 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     1801 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     1861 gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta cggcacctcg
     1921 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     1981 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     2041 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttgccgattt
     2101 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     2161 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     2221 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     2281 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     2341 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     2401 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     2461 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     2521 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     2581 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     2641 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     2701 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     2761 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     2821 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     2881 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     2941 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     3001 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     3061 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     3121 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     3181 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     3241 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     3301 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     3361 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     3421 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     3481 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     3541 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3601 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     3661 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     3721 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     3781 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     3841 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     3901 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     3961 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     4021 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     4081 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     4141 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     4201 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     4261 ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt aggtatctca
     4321 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     4381 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     4441 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     4501 cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta tttggtatct
     4561 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     4621 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     4681 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     4741 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     4801 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     4861 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     4921 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     4981 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     5041 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     5101 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     5161 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     5221 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     5281 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     5341 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     5401 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     5461 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     5521 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     5581 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     5641 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     5701 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     5761 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     5821 ttccgcgcac atttccccga aaagtgccac ctgacgtc
//
