LOCUS       Exported                7687 bp ds-DNA     circular SYN 19-NOV-2023
DEFINITION  260_TSS20231103-0571-01100.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7687)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Nov 6, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7687
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        complement(66..84)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     polyA_signal    110..334
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      380..808
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        822..1151
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1002..1137
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1218..2012
                     /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"
     polyA_signal    2186..2307
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2356..2372)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2380..2396
                     /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(2404..2434)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2449..2470
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     misc_feature    2576..3516
                     /label=ColE1
                     /note="ColE1"
     rep_origin      complement(2758..3346)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(3517..4377)
                     /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"
     promoter        complement(4378..4482)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4748..5127
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5128..5331
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     promoter        5376..5394
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      5414..5423
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             5420..6121
                     /codon_start=1
                     /label=mBaoJin
                     /translation="MVSKGEEENMASTPFKFQLKGTINGKSFTVEGEGEGNSHEGSHKG
                     KYVCTSGKLPMSWAALGTTFGYGMKYYTKYPSGLKNWFREVMPGGFTYDRHIQYKGDGS
                     IHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTGDMNVSLPNEVPQIPRDDGVECPVTL
                     LYPLLSDKSKYVEAHQYTICKPLHNQPAPDVPYHWIRKQYTQSKDDAEERDHICQSETL
                     EAHLKGMDELYK"
     CDS             6182..7537
                     /codon_start=1
                     /label=TUBA1B
                     /translation="VRECISIHVGQAGVQIGNACWELYCLEHGIQPDGQMPSDKTIGGG
                     DDSFNTFFSETGAGKHVPRAVFVDLEPTVIDEVRTGTYRQLFHPEQLITGKEDAANNYA
                     RGHYTIGKEIIDLVLDRIRKLADQCTGLQGFLVFHSFGGGTGSGFTSLLMERLSVDYGK
                     KSKLEFSIYPAPQVSTAVVEPYNSILTTHTTLEHSDCAFMVDNEAIYDICRRNLDIERP
                     TYTNLNRLISQIVSSITASLRFDGALNVDLTEFQTNLVPYPRIHFPLATYAPVISAEKA
                     YHEQLSVAEITNACFEPANQMVKCDPRHGKYMACCLLYRGDVVPKDVNAAIATIKTKRS
                     IQFVDWCPTGFKVGINYQPPTVVPGGDLAKVQRAVCMLSNTTAIAEAWARLDHKFDLMY
                     AKRAFVHWYVGEGMEEGEFSEAREDMAALEKDYEEVGVDSVEGEGEEEGEEY"
     promoter        complement(7565..7583)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
ORIGIN
        1 gttgaggaga gggtgaggaa gaaggagagg aatactaagg atccaccgga tctagagggc
       61 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
      121 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
      181 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
      241 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
      301 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
      361 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
      421 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
      481 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
      541 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
      601 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
      661 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
      721 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
      781 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
      841 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
      901 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
      961 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     1021 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     1081 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     1141 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cggatctgat caagagacag
     1201 gatgaggatc gtttcgcatg attgaacaag atggattgca cgcaggttct ccggccgctt
     1261 gggtggagag gctattcggc tatgactggg cacaacagac aatcggctgc tctgatgccg
     1321 ccgtgttccg gctgtcagcg caggggcgcc cggttctttt tgtcaagacc gacctgtccg
     1381 gtgccctgaa tgaactgcag gacgaggcag cgcggctatc gtggctggcc acgacgggcg
     1441 ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg aagggactgg ctgctattgg
     1501 gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc tcctgccgag aaagtatcca
     1561 tcatggctga tgcaatgcgg cggctgcata cgcttgatcc ggctacctgc ccattcgacc
     1621 accaagcgaa acatcgcatc gagcgagcac gtactcggat ggaagccggt cttgtcgatc
     1681 aggatgatct ggacgaagag catcaggggc tcgcgccagc cgaactgttc gccaggctca
     1741 aggcgcgcat gcccgacggc gaggatctcg tcgtgaccca tggcgatgcc tgcttgccga
     1801 atatcatggt ggaaaatggc cgcttttctg gattcatcga ctgtggccgg ctgggtgtgg
     1861 cggaccgcta tcaggacata gcgttggcta cccgtgatat tgctgaagag cttggcggcg
     1921 aatgggctga ccgcttcctc gtgctttacg gtatcgccgc tcccgattcg cagcgcatcg
     1981 ccttctatcg ccttcttgac gagttcttct gagcgggact ctggggttcg aaatgaccga
     2041 ccaagcgacg cccaacctgc catcacgaga tttcgattcc accgccgcct tctatgaaag
     2101 gttgggcttc ggaatcgttt tccgggacgc cggctggatg atcctccagc gcggggatct
     2161 catgctggag ttcttcgccc accccaactt gtttattgca gcttataatg gttacaaata
     2221 aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt ctagttgtgg
     2281 tttgtccaaa ctcatcaatg tatcttatca tgtctgtata ccgtcgacct ctagctagag
     2341 cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc
     2401 acacaacata cgagccggaa gcataaagtg taaagcctgg ggtgcctaat gagtgagcta
     2461 actcacatta attgcgttgc gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca
     2521 gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc
     2581 cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc
     2641 tcactcaaag gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat
     2701 gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt
     2761 ccataggctc cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg
     2821 aaacccgaca ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc
     2881 tcctgttccg accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt
     2941 ggcgctttct catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa
     3001 gctgggctgt gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta
     3061 tcgtcttgag tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa
     3121 caggattagc agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa
     3181 ctacggctac actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt
     3241 cggaaaaaga gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt
     3301 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     3361 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     3421 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     3481 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     3541 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     3601 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     3661 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     3721 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     3781 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     3841 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     3901 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     3961 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     4021 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     4081 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     4141 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     4201 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     4261 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     4321 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     4381 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     4441 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     4501 gccacctgac gtcgacggat cgggagatct cccgatcccc tatggtgcac tctcagtaca
     4561 atctgctctg atgccgcata gttaagccag tatctgctcc ctgcttgtgt gttggaggtc
     4621 gctgagtagt gcgcgagcaa aatttaagct acaacaaggc aaggcttgac cgacaattgc
     4681 atgaagaatc tgcttagggt taggcgtttt gcgctgcttc gcgatgtacg ggccagatat
     4741 acgcgttgac attgattatt gactagttat taatagtaat caattacggg gtcattagtt
     4801 catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc gcctggctga
     4861 ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat agtaacgcca
     4921 atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc ccacttggca
     4981 gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga cggtaaatgg
     5041 cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg gcagtacatc
     5101 tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat caatgggcgt
     5161 ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt caatgggagt
     5221 ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc cgccccattg
     5281 acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc tctctggcta
     5341 actagagaac ccactgctta ctggcttatc gaaattaata cgactcacta tagggagacc
     5401 caagcttggt accgccacca tggtctcaaa gggagaggag gaaaacatgg ctagtacacc
     5461 atttaaattt caacttaaag gaaccatcaa tggcaaatcg tttaccgttg aaggcgaagg
     5521 tgaagggaac tcacatgaag gttctcataa aggaaaatat gtttgtacaa gtggaaaact
     5581 accgatgtca tgggcagcac ttgggacaac ctttggttat ggaatgaaat attataccaa
     5641 atatcctagt ggactgaaga actggtttcg tgaagtaatg cccggaggct ttacctacga
     5701 tcgtcatatt caatataaag gcgatgggag tatccatgca aaacaccaac actttatgaa
     5761 aaatgggact tatcacaaca ttgtagaatt taccggtcag gattttaaag aaaatagtcc
     5821 agtcttaact ggagatatga atgtctcatt accgaatgaa gtcccacaaa tacccagaga
     5881 tgatggagta gaatgcccag tgaccttgct ttatccttta ttatcggata aatcaaaata
     5941 cgtcgaggct caccaatata caatctgcaa gcctcttcat aatcaaccag cacctgatgt
     6001 cccatatcac tggattcgta aacaatacac acaaagcaaa gatgatgccg aggaacgcga
     6061 tcatatctgt caatcagaga ctctcgaagc acacttaaag ggcatggacg agctgtataa
     6121 gggaggtggt ggatcaggag gaggtggttc tggaggtggt ggatcaggag gaagatctcg
     6181 agtgcgtgag tgcatctcca tccacgttgg ccaggctggt gtccagattg gcaatgcctg
     6241 ctgggagctc tactgcctgg aacacggcat ccagcccgat ggccagatgc caagtgacaa
     6301 gaccattggg ggaggagatg actccttcaa caccttcttc agtgagacgg gcgctggcaa
     6361 gcacgtgccc cgggctgtgt ttgtagactt ggaacccaca gtcattgatg aagttcgcac
     6421 tggcacctac cgccagctct tccaccctga gcagctcatc acaggcaagg aagatgctgc
     6481 caataactat gcccgagggc actacaccat tggcaaggag atcattgacc ttgtgttgga
     6541 ccgaattcgc aagctggctg accagtgcac cggtcttcag ggcttcttgg ttttccacag
     6601 ctttggtggg ggaactggtt ctgggttcac ctccctgctc atggaacgtc tctcagttga
     6661 ttatggcaag aagtccaagc tggagttctc catttaccca gcaccccagg tttccacagc
     6721 tgtagttgag ccctacaact ccatcctcac cacccacacc accctggagc actctgattg
     6781 tgccttcatg gtagacaatg aggccatcta tgacatctgt cgtagaaacc tcgatatcga
     6841 gcgcccaacc tacactaacc ttaaccgcct tattagccag attgtgtcct ccatcactgc
     6901 ttccctgaga tttgatggag ccctgaatgt tgacctgaca gaattccaga ccaacctggt
     6961 gccctacccc cgcatccact tccctctggc cacatatgcc cctgtcatct ctgctgagaa
     7021 agcctaccat gaacagcttt ctgtagcaga gatcaccaat gcttgctttg agccagccaa
     7081 ccagatggtg aaatgtgacc ctcgccatgg taaatacatg gcttgctgcc tgttgtaccg
     7141 tggtgacgtg gttcccaaag atgtcaatgc tgccattgcc accatcaaaa ccaagcgcag
     7201 catccagttt gtggattggt gccccactgg cttcaaggtt ggcatcaact accagcctcc
     7261 cactgtggtg cctggtggag acctggccaa ggtacagaga gctgtgtgca tgctgagcaa
     7321 caccacagcc attgctgagg cctgggctcg cctggaccac aagtttgacc tgatgtatgc
     7381 caagcgtgcc tttgttcact ggtacgtggg tgaggggatg gaggaaggcg agttttcaga
     7441 ggcccgtgaa gatatggctg cccttgagaa ggattatgag gaggttggtg tggattctgt
     7501 tgaaggagag ggtgaggaag aaggagagga atactaagga tccaccggat ctagagggcc
     7561 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     7621 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     7681 ctcccac
//
