LOCUS       158_pBlueScript         4967 bp    DNA     circular SYN 15-JAN-2025
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4967)
  AUTHORS   NYU School of Medicine
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2025 from SnapGene 8.0.1
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4967
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(3..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        626..644
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    680..1001
                     /label=5'HA Podxl
     CDS             1018..1071
                     /codon_start=1
                     /product="2A peptide from Thosea asigna virus capsid
                     protein"
                     /label=T2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             join(1072..1074,1075..1785)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKV
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLE
                     FVTAAGITLGMDELYK"
     polyA_signal    1819..1940
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    1941..2742
                     /label=3'HA Podxl
     promoter        complement(2779..2797)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(2818..2834)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    2842..2858
                     /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(2866..2872,2873..2890,2891..2896))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2911..2932
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(3220..3808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(join(3979..4770,4771..4839))
                     /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(4840..4944)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgcgtaata cgactcacta tagggcgaat tgggtaccgg
      661 gccccccctc gaggtcgaca gtcttcgcag caacgtgcgc ggggccgcag cggcgcagac
      721 accgcccggg ccgcagacac cttcgctcgt cgctgctggc tccgtgtccc agcccagctc
      781 tgcggtaggc aggaggagcg gccggatcgc gactccggag ttcgcggccg tcgccaggcc
      841 tcccggagcg gcgccgacgc tcccgctgtc cggacgcgcg accctgcgcc agcgccgcag
      901 ccacctgctc cgagtccccg ggcgagcaca accatgcctc ccactacggc gctctccgcg
      961 ctgctgctgc tgctactgtc gcctgcatct cactcccata atgaattcgg cagtggagag
     1021 ggcagaggaa gtctgctaac atgcggtgac gtcgaggaga atcctggccc agtgagcaag
     1081 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
     1141 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
     1201 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
     1261 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
     1321 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
     1381 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1441 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1501 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1561 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1621 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1681 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1741 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaaggagtt ctaactagag
     1801 ctcgctgatc agcctcgaaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     1861 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     1921 aaactcatca atgtatctta ggtgagtgga cagccgcggc tctgccggga cccctggtca
     1981 ttcccctccg ccccaagcct ggtggggctc ctgggaagcc ggctgcaccg cgtggtcgga
     2041 ccgcttgcag cctctcccgc gtggccgcac gcagagtcct gaccttggca ccatccacgc
     2101 cccctccaaa ttggagaacc ccgcaggtgc ggggccagat cgtggattta ttcctagtct
     2161 tcactggtgt agagtgagcc ggaggttgtg agattgtaaa gggggcactt ggtgggttcg
     2221 ctggagatgg gggggaaact ttgggttgtg gggaggtccc agattcccgc atgaccacca
     2281 gaatgcgatt tctctttctc gtgtttgggg ctactagggg accgtattgg gaggtcgttg
     2341 aaactaagca aagccgcggg cagggaccac aagaaggagc ccctgaaact gacctcaaat
     2401 ccctaaatgc ccacgcgtgt agcgtacttc ttgtgcaagc gctttttctt aagcttgtcc
     2461 cgggacgggt ggaataagcg ctccttccag ggtcccagag cctggttcag tggggtcctt
     2521 gccttcatgt tgccaaagga ccggattctt tctcagagca tggccaggat ggtgggctcc
     2581 tcccacccct ttgtgtaaac aagaaatttc tttcacaaat gaggtttgag ttgtgtctta
     2641 gtaatacctc gggcacccat ctcccatcag acccagcttg gagacgatcc cactaccttc
     2701 tttctgaagg tggaatgtta ctttctgttt gagaggtcag gcgcggccgc caccgcggtg
     2761 gagctccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt aatcatggtc
     2821 atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg
     2881 aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt
     2941 gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg
     3001 ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga
     3061 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     3121 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     3181 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     3241 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     3301 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     3361 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     3421 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     3481 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     3541 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     3601 gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag
     3661 gacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     3721 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     3781 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     3841 cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat
     3901 cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga
     3961 gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg
     4021 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     4081 gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc
     4141 agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac
     4201 tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc
     4261 agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc
     4321 gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc
     4381 catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt
     4441 ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc
     4501 atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg
     4561 tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag
     4621 cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat
     4681 cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc
     4741 atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa
     4801 aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta
     4861 ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa
     4921 aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccac
//
