LOCUS       Exported                6197 bp ds-DNA     circular SYN 10-FEB-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6197)
  AUTHORS   jay
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Apr 24, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6197
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        378..1556
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          609..1547
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     promoter        1623..1641
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1677..1839
                     /label=QBI SP163
                     /note="strong translational enhancer derived from the 5' 
                     UTR of the mouse VEGF gene"
     CDS             1840..2136
                     /codon_start=1
                     /label=NRL-205 (truncated)
                     /translation="MSVRELNRQLRGCGRDEALRLKQRRRTLKNRGYAQACRSKRLQQR
                     RGLEAERARLAAQLDALRAEVARLARERDLYKARCDRLTSSGPGSGDPSHLFL"
     polyA_signal    2210..2434
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2480..2908
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        2922..3252
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3103..3238
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        3300..3347
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3366..3740
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    3870..3991
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4040..4056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4064..4080
                     /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(4088..4118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4133..4154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(4442..5030)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5201..6061)
                     /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(6062..6166)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtcgactct 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 tagcaaattg ggacattgat tattgactag gcttttgcaa aaagctttgc
      301 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      361 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      421 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      481 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      541 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      601 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      661 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
      721 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
      781 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
      841 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
      901 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
      961 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1021 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1081 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1141 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1201 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1261 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1321 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1381 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1441 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1501 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa
     1561 ttagcttggt accgtcctct ctggctaact agagaaccca ctgcttactg gcttatcgaa
     1621 attaatacga ctcactatag ggagacccaa gctggctagc gtttaaactt aagcttagcg
     1681 cagaggcttg gggcagccga gcggcagcca ggccccggcc cgggcctcgg ttccagaagg
     1741 gagaggagcc cgccaaggcg cgcaagagag cgggctgcct cgcagtccga gccggagagg
     1801 gagcgcgagc cgcgccggcc ccggacggcc tccgaaacca tgtctgtgcg ggagctaaac
     1861 cggcagctgc ggggctgcgg gcgcgacgag gcgctgcggc tgaagcagag gcgccgcacg
     1921 ctgaagaacc gcggctacgc gcaggcctgt cgctccaagc ggctgcagca gcggcgcggg
     1981 ctggaggccg agcgcgcccg cctggccgcc cagctggacg cgctgcgggc cgaggtggcc
     2041 cgcctggccc gggagcgcga tctctacaag gctcgctgtg accggctaac ctcgagcggc
     2101 cccgggtccg gggacccctc ccacctcttc ctctgagccg ttcagagcac cttgtggtgg
     2161 cggccgctcg agtctagagg gcccgtttaa acccgctgat cagcctcgac tgtgccttct
     2221 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     2281 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     2341 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     2401 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     2461 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2521 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2581 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcggggcatc
     2641 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2701 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2761 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2821 gtctattctt ttgatttata agggattttg gggatttcgg cctattggtt aaaaaatgag
     2881 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2941 gaaagtcccc aggctcccca ggcaggcaga agtatgcaaa gcatgcatct caattagtca
     3001 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     3061 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     3121 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     3181 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     3241 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     3301 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     3361 aaaccatggc caagttgacc agtgccgttc cggtgctcac cgcgcgcgac gtcgccggag
     3421 cggtcgagtt ctggaccgac cggctcgggt tctcccggga cttcgtggag gacgacttcg
     3481 ccggtgtggt ccgggacgac gtgaccctgt tcatcagcgc ggtccaggac caggtggtgc
     3541 cggacaacac cctggcctgg gtgtgggtgc gcggcctgga cgagctgtac gccgagtggt
     3601 cggaggtcgt gtccacgaac ttccgggacg cctccgggcc ggccatgacc gagatcggcg
     3661 agcagccgtg ggggcgggag ttcgccctgc gcgacccggc cggcaactgc gtgcacttcg
     3721 tggccgagga gcaggactga cacgtgctac gagatttcga ttccaccgcc gccttctatg
     3781 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg
     3841 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca
     3901 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     3961 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct
     4021 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     4081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     4141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     4201 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct
     4261 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     4321 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     4381 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     4441 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     4501 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     4561 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     4621 gcgtggcgct ttctcaatgc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     4681 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     4741 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     4801 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     4861 ctaactacgg ctacactaga aggacagtat ttggtatctg cgctctgctg aagccagtta
     4921 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     4981 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5041 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5101 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     5161 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     5221 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     5281 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     5341 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     5401 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     5461 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     5521 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     5581 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5641 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5701 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5761 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5821 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5881 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5941 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6001 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6061 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6121 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     6181 aagtgccacc tgacgtc
//
